1. science
2. chemistry

Sample
Preparation
Nucleic Acid Sample Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
RNA Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
DNA Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
DNA Cleanup and Purification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Protein Sample Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
Protein Sample Preparation Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Protein Sample Preparation Instruments . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Protein Sample Preparation Cartridges and Columns . . . . . . . . . . . . . . . . . . 19
sample preparation
nucleic acid sample preparation
Nucleic Acid Sample Preparation
Bio-Rad offers a broad range of sample preparation products for nucleic acid research. These products offer efficiency
and throughput in both small- and large-scale purification of DNA, RNA, and PCR products with reproducible yield
and purity. For more information, go to www.bio-rad.com/nasampleprep/
RNA Isolation
see also
Amplification reagents:
pages 389–400.
A variety of products enable isolation of total RNA that is highly pure, intact, and suitable for a variety of downstream
applications, including quantitative real-time PCR, northern blotting, and microarray analysis:
Real-time PCR
systems:
pages 383–389.
• Aurum™
total RNA membrane-based kits — these kits deliver DNA-free total RNA for sensitive applications in
both 96-well plate and mini spin-column formats for streamlined RNA isolation
Microarray products:
pages 342–344.
•PureZOL™
RNA isolation reagent — a monophasic solution of phenol and guanidine isothiocyanate, an improved
version of the single-step total RNA isolation method developed by Chomczynski and Sacchi (1987)
RNA Isolation Selection Guide
Aurum Total RNA Kits
Mini
Fatty and Fibrous Tissue
96
Format
Mini column
Filtration (vacuum or spin)
Mini column
Filtration (vacuum or spin)
96-well plate
Filtration (vacuum or spin)
—
Maximum starting
amounts
Cultured cells
2 x 106
—
Bacterial cells
2.4 x 109
—
Yeast cells3 x 107
—
Hard animal tissue 20 mg100 mg
Soft to moderately 40 mg100 mg hard animal tissue
Plant tissue
40 mg100 mg
Isolation method
Silica membrane Lysis with PureZOL reagent, purification on silica membrane
Number of preps10* or 50 mini preps10* or 50 mini preps 1 x 106
1 x 107
8 x 108
2.4 x 109
2 x 107
3 x 107
—100 mg
—100 mg
—100 mg
Silica membrane
Organic extraction
2 x 96-well plate preps
50 or 100 preps (1 ml/prep)
Number of washes333
—
DNase I included**
No
Yes
Yes
PureZOL RNA
Isolation Reagent
Yes
DNase I digest time15 min (animal tissue, 25 min)15 min10 min
—
Total preparation time <50 min (1.5 hr with DNase I)
<50 min (1.5 hr with DNase I)
<60 min
<60 min
—
Binding capacity
>100 µg
>100 µg >40 µg
Elution volume
2 x 40 µl
2 x 40 µl
80 µl30–100 µl
Yield***
28 µg/2 x 106 NIH 3T3 cells
96 µg/100 mg brain10 µg/1 x 106 NIH 3T3 cells1.15 µg/mg brain
38 µg/2 x 106 HeLa cells
58 µg/100 mg breast17 µg/1 x 106 HeLa cells1.37 µg/mg fat
20 µg/40 mg brain
85 µg/100 mg heart11 µg/2 x 107 S. cerevisiae cells1.2 µg/mg heart
52 µg/20 mg thymus
63 µg/100 mg skin
5 µg/8 x 108 E. coli cells
0.66 µg/mg cartilage
132 µg/40 mg spleen
54 µg/100 mg cartilage
5 µg/8 x 108 B. cereus cells1.2 µg/mg skin
36 µg/40 mg lung1.0 µg/mg potato
32 µg/40 mg potato1.37 µg/1 x 105 293H cells
28 µg/3 x 107 S. cerevisiae cells 0.1 µg/mg Arabidopsis
54 µg/2.4 x 109 E. coli cells
0.38 µg/mg A. niger
0.63 µg/2.4 x 107
E. coli cells
0.31 µg/3 x 105
S. cerevisiae cells
Purity
A260/A280 of 1.9–2.1
A260/A280 of 1.9–2.1
A260/A280 of 1.9–2.1
*Trial sizes of the Aurum total RNA mini kit and Aurum total RNA fatty and fibrous tissue kit are available.
** Removal not required.
*** Yields will vary depending on the developmental stage, tissue type or cell line, and growth conditions used.
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A260/A280 of 1.9–2.1
sample preparation
nucleic acid sample preparation
Aurum™ Total RNA Kits
Aurum total RNA kits produce high-quality RNA for all major applications. After sample collection and enzymatic
disruption (if necessary), the sample is homogenized in a lysis buffer. The lysate is mixed with alcohol, loaded onto
a silica membrane in a spin column or a 96-well plate, then washed and digested on-column or in-well with
DNase I to remove genomic DNA. Once purified RNA is eluted from the membrane, it is ready for use without further
manipulation. Aurum total RNA kits:
• Isolate
high yields of RNA from a wide range of sample types
RNA that is DNA- and phenol-free
• Produce ready-to-use RNA suitable for the most demanding downstream applications
• Use a quick, convenient protocol
• Isolate RNA using either a centrifuge or a vacuum manifold
• Include lyophilized DNase I
• Provide RNase-free reagents and plasticware
see also
PureZOL reagent:
page 5.
Transfection reagents:
pages 351–352.
Nucleic acid
amplification:
pages 374–412.
• Purify
Aurum total RNA kits are designed for streamlined processing using a choice of vacuum- or spin-mediated
protocols. If vacuum-mediated purification is preferred, the protocol may be carried out on the Aurum vacuum
manifold (page 4), which can process either a 96-well plate or 18 spin columns at one time.
Aurum Total RNA Mini Kit
The Aurum total RNA mini kit produces DNA-free total
RNA from a wide range of starting materials, including
cultured cells, bacteria, and yeast, as well as animal and
plant tissues. The kit utilizes a stringent reagent composed
of guanidine isothiocyanate and b-mercaptoethanol
for efficient sample lysis and quick RNase inactivation,
followed by purification on silica membrane in a spincolumn format using a spin- or vacuum-mediated
protocol. The Aurum total RNA mini kit can also be used
for RNA cleanup and desalting.
For more information, request bulletin 2920.
Aurum Total RNA Fatty and Fibrous Tissue Kit
The Aurum total RNA fatty and fibrous tissue kit produces
total RNA from samples that are difficult to disrupt. The
kit is perfect for purifications involving fatty and fibrous
tissues or samples such as fungi that are rich in RNases.
The kit utilizes PureZOL™ RNA reagent, a potent phenolbased reagent, which effectively lyses tissues and cells.
The kit includes mini spin columns that can be processed
using a choice of vacuum- or spin-mediated protocols.
For more information, request bulletin 5282.
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sample preparation
nucleic acid sample preparation
Aurum Total RNA 96 Kit
The Aurum total RNA 96 kit reproducibly isolates DNA-free total RNA from
cultured cells, bacteria, and yeast in a 96-well format for high-throughput total RNA
isolation. The total RNA binding plate is designed for use on the Aurum vacuum
manifold (below). The entire purification process is completed in under 60 minutes.
For more information, request bulletin 2919.
Aurum Vacuum Manifold
Vacuum-mediated RNA purifications can be carried out in the Aurum vacuum
manifold. The manifold serves as a vacuum purification platform for either
96-well plates or individual luer-ended spin columns. Benefits include:
• Streamlined
processing of up to 18 mini columns at one time
•Universal
manifold that converts from plate to column format
by inserting the column adaptor plate
Consumables and Accessories
The reagents and plasticware used in the Aurum RNA kits are available as refill orders. These products can be ordered
separately from the Aurum total RNA kits.
Ordering Information
Catalog #
Description
Price, US $
Aurum Total RNA Mini Kit
732-6820
Aurum Total RNA Mini Kit, 50 preps, includes 50 RNA binding columns, 50 capless collection tubes (2.0 ml), 165.00
2 x 50 capped sample tubes (2.0 ml), 50 capped sample tubes (1.5 ml), 1 vial lyophilized DNase I,
RNase-free reagents, protocol overview, instructions
Aurum Total RNA Fatty and Fibrous Tissue Kit
732-6830
Aurum Total RNA Fatty and Fibrous Tissue Kit, 50 preps, includes 732-6870, 50 ml PureZOL RNA isolation reagent 270.00
Aurum Total RNA Fatty and Fibrous Tissue Module, 50 preps, includes 50 RNA binding columns, 182.00
732-6870*
50 capless collection tubes (2.0 ml), 100 capped sample tubes (2.0 ml), 50 capped sample tubes (1.5 ml), 1 ml of lyophilized DNase I, RNase-free reagents and plasticware, instructions
Aurum Total RNA 96 Kit
Aurum Total RNA 96 Kit, 2 x 96-well preps, includes 2 grow blocks, 2 growth membranes, sealing tape, 2 RNA 732-6800
binding plates, 2 collection microplates, 2 vials lyophilized DNase I, RNase-free reagents, protocol overview, instructions
441.00
Aurum Vacuum Manifold
732-6470
Aurum Vacuum Manifold, includes column adaptor plate, 4 replacement luer caps, A stage and B stage, waste collection tray, vacuum regulator and gauge, tubing, protocol overview, instructions
562.00
Consumables
732-6890
732-6880
732-6828
732-6801
732-6802
732-6803
732-6804
732-6805
732-6826
and Accessories
PureZOL RNA Isolation Reagent, 100 ml139.00
PureZOL RNA Isolation Reagent, 50 ml
83.00
DNase I, RNase-free, lyophilized, 1 vial
45.00
Aurum Total RNA Elution Solution, RNase-free, 20 ml13.00
Aurum Total RNA Lysis Solution, RNase-free, 85 ml39.00
Aurum Total RNA Wash High-Stringency Solution, RNase-free, 150 ml32.00
Aurum Total RNA Wash Low-Stringency Solution, 60 ml
24.00
Aurum DNase Dilution Solution, 20 ml11.00
Aurum RNA Binding Mini Columns, 50
27.00
* Not provided with PureZOL RNA isolation reagent (see 732-6890 or 732-6880 to order separately).
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sample preparation
nucleic acid sample preparation
PureZOL™ RNA Isolation Reagent
see also
The PureZOL RNA isolation reagent protocol is an improvement over the rapid, widely
used, and proven method of RNA isolation developed by Chomczynski and Sacchi
(1987). PureZOL RNA isolation reagent is a potent monophasic combination of phenol
and the chaotropic agent guanidine isothiocyanate, which effectively lyses cells and
tissues, deproteinates RNA, and inactivates endogenous nucleases in a single step.
DNA and protein are efficiently removed from the RNA following phase separation.
Aurum total RNA kits:
pages 3–4.
The ready-to-use PureZOL RNA isolation reagent is a versatile and efficient means
of isolating high yields of RNA from a variety of sources, including cultured cell, animal and
plant tissue, yeast, virus, and bacteria samples. The single-solution format permits recovery
of RNA from small quantities of tissues or cells, making it ideally suited for gene expression studies or whenever limited
sample quantities are available. Total RNA isolated using PureZOL is free of DNA and protein and can be used for northern
blot analysis, in vitro translation, poly(A)+ selection, RNase protection assays, RT-PCR, and molecular cloning. Because no
spin columns are used, protocols are scalable to accommodate a wide range of sample sizes.
Ordering Information
Catalog #
732-6880
732-6890
Description
Price, US $
PureZOL RNA Isolation Reagent, 50 ml
83.00
PureZOL RNA Isolation Reagent, 100 ml139.00
DNA Isolation
Obtaining pure, intact DNA is essential to the quality of downstream analysis techniques, including automated
fluorescent sequencing, cloning, transformation, and cell culture transfection. Bio-Rad offers plasmid and Chelex®based extraction products for a variety of starting sample types.
• Aurum™
plasmid mini kit — silica membrane-based technology and optimized reagents for binding more than
20 µg of plasmid DNA with excellent purity and the ease of either vacuum or spin formats
see also
Competent cells:
page 96.
Transfection reagents:
pages 351–352.
Nucleic acid
amplification:
pages 374–412.
Prep® kits — matrix-based plasmid preps with A260/A280 values of 1.75–1.85 and >99% sequencing
accuracy over 650 bases on an ABI PRISM 3700 fluorescent capillary sequencer
• Quantum
Plasmid Purification Kit Selection Guide
Kit
DNA Yield
Preparation Time
Bacterial Culture Density or Volume
Binding Matrix
Growth Media Silica membrane
Standard
Quantum Prep® >20 µg15 min1–10 ml Spin
miniprep
Diatomaceous
earth
Enriched
or standard
Quantum Prep >300 µg
45 min
20–40 ml
Spin
midiprep
Diatomaceous earth
Enriched
or standard
Aurum™ plasmid >20 µg10 min
Up to 12 OD • ml* mini
Purification Format
Vacuum**
and spin
* (OD600 of undiluted culture) x (culture volume in ml) = # of OD • ml. 1 OD600 is equivalent to approximately 8 x 108 cells/ml.
** Using the Aurum vacuum manifold.
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sample preparation
nucleic acid sample preparation
Aurum™ Plasmid Mini Kit
The easy-to-use Aurum plasmid mini kit improves the efficiency
and throughput of plasmid purifications with a simple bind-washelute protocol using silica membranes. Lysed bacterial cultures
can be cleared either by centrifugation or by vacuum filtration with
the Aurum vacuum manifold. The clarified lysates are then applied
to plasmid binding columns and plates, where DNA is bound, then
washed, and finally eluted — all in <10 minutes. The purified plasmid
DNA can be immediately used in any downstream molecular biology application.
Applications
Aurum plasmid mini kits deliver high yields of reproducible plasmid DNA preparations for:
• Automated
• Transfection
• Restriction
• PCR
fluorescent sequencing
digestion
• Ligation and transformation
For more information, request bulletin 2664.
Ordering Information
Catalog #
732-6400 732-6470 Description
Price, US $
Aurum Plasmid Mini Kit, 100 preps, includes plasmid-binding mini columns, 100 capless collection tubes, 140.00
reagents, protocol overview, instructions
Aurum Vacuum Manifold, includes column adaptor plate, 4 replacement luer caps, A stage and B stage, 562.00
waste collection tray, vacuum regulator and gauge, tubing, protocol overview, instructions
Quantum Prep® Plasmid Kits
The consistent high yields achieved with Quantum Prep plasmid kits are due to the unique Quantum Prep DNA
binding matrix. This patented* matrix, a special grade of diatomaceous earth, is available only from Bio-Rad. Its large
surface area enables the use of enriched growth media such as Terrific Broth to obtain yields in milligram quantities.
All Quantum Prep kits use an easy spin-based procedure to quickly go from cell culture to purified DNA.
Plasmid purification with the Quantum Prep kits uses an optimized
alkaline lysis procedure and takes advantage of the high binding affinity
of DNA to the Quantum Prep binding matrix. Bacterial cultures are
spin-clarified after lysis and neutralization. The matrix is then added to
the clarified lysate to enhance binding of plasmid DNA. Slurried matrix
containing bound DNA is added to the columns for washing and elution
by low-speed centrifugation. The DNA is suitable for use in:
• Automated
fluorescent sequencing
• Restriction digestion
• Ligation and transformation
• Transfection
• PCR
Quantum Prep Miniprep Kit
Quantum Prep miniprep kits offer high quality and yield in an easy
spin-based procedure that takes less than 15 minutes from cell culture
to purified plasmid. DNA is recovered in water or TE for immediate use
in all downstream molecular biology applications.
* U.S. patent 5,075,430.
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Quantum Prep kit
Brand X
Brand Y
Phenol/chloroform
0
51015
20
25
Plasmid yield in µg/1.5 ml culture
High yield of Quantum Prep kits. Yields as high
as 24 µg can be obtained from a 1.5 ml culture of
DH5aF'[pB1] grown in Terrific Broth.
sample preparation
nucleic acid sample preparation
Quantum Prep Plasmid Midiprep Kit
Quantum Prep plasmid midiprep kit uses a simple spin-column
procedure that significantly reduces the time required to generate the
large quantities of plasmid DNA needed to support applications such
as transfection, subcloning, and other enzymatic manipulations.
For more information, request bulletin 2325.
Ordering Information
Catalog #
732-6100
732-6120
Description
Price, US $
Quantum Prep Plasmid Miniprep Kit, 100 preps, includes 20 ml cell resuspension solution, 25 ml cell 125.00
lysis solution, 25 ml neutralization solution, 20 ml Quantum Prep matrix, 63 ml wash solution, 100 spin columns, instructions
Quantum Prep Plasmid Midiprep Kit, 20 preps, includes 110 ml cell resuspension solution, 110 ml 125.00
cell lysis solution, 110 ml neutralization solution, 20 ml Quantum Prep matrix, 125 ml wash solution, 20 midi
spin columns, instructions
Chelex®-Based Resins
Two Chelex-based resins are available for DNA extraction from a variety of starting samples such as blood, bacteria,
tissue, and hair. Because extracted DNA is commonly used for downstream PCR, it must be free of metal ions
that could inhibit the amplification process. Chelex is composed of paired iminodiacetate ions coupled to a styrene
divinylbenzene matrix, providing high selectivity for polyvalent cations and effective removal of these contaminants.
see also
Molecular biology
and biotechnology
grade resins:
pages 45–46.
Chelex 100 Molecular Biology Grade Resin
Unlike other Chelex-based DNA extraction resins, Chelex 100 (200–400 mesh) molecular biology grade resin
is a highly pure nuclease- and ligase inhibitor-free chelating resin certified not to interfere with PCR. This pure,
pipettable, small-scale resin ensures complete removal of PCR inhibitors and contaminating metal ions.
InstaGene™ Matrix
InstaGene matrix is designed for the rapid isolation of small amounts of genomic DNA of sufficient purity for PCR. The
specially formulated 6% w/v Chelex resin adsorbs cell lysis products that interfere with PCR, leaving genomic DNA
template in the supernatant, where it is immediately available for PCR reactions. Although intended specifically for PCR
applications, the InstaGene matrix eliminates the deproteinization, organic extraction, dialysis, and alcohol precipitation
steps required in traditional genomic DNA purification procedures. Features include:
•Ease
of use — the small-particle-size slurry won’t clog pipet tips and is easily resuspended with the included stirbar
matrix concentration ready upon resuspension
• Affordability and cost-effectiveness — 100 samples from 1 bottle
• Appropriate
For more information, request bulletin 2074.
InstaGene Protocol
PCR-ready
Add bacterial colony
to water.
Spin and remove
supernatant.
Add InstaGene matrix and incubate at 56°C for
30 min. Vortex and incubate at 100°C for 8 min.
Spin; use
supernatant for PCR.
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sample preparation
nucleic acid sample preparation
Ordering Information
Catalog #
142-1253
732-6030
Description
Price, US $
Chelex 100 Resin, molecular biology grade, 50 g, sodium, 200–400 dry mesh, 75–150 µm wet bead143.00
InstaGene Matrix, 20 ml, enough for 100 samples
43.00
DNA Cleanup and Purification
Bio-Rad offers a variety of pre-PCR and post-PCR sample preparation and cleanup columns and prepacked columns; many
are also suitable for DNA fragment purification. Refer to the guide below to select the most suitable products for your needs.
PCR Product and DNA Fragment Purification Guide
Freeze ’N Squeeze™ DNA Gel Extraction Spin Columns
Micro Bio-Spin™ and
Bio-Spin® Columns
PCR Kleen™ Spin Columns Application
Recovery of DNA fragments (50 bp–23 kb) purified from TAE- or TBE-buffered agarose gels
Dye terminator and label cleanup
from sequencing reactions; buffer exchange; desalting
Removal of PCR products
>200 bp from other reaction
mixture components
Starting material
≤700 mg gel slice10–100 µl of sequencing reaction mixture
25–100 µl of PCR or other
enzyme reaction
Format
Spin-column extraction of frozen-thawed gel slices
Prepacked size exclusion
spin column
Prepacked size exclusion
spin column
Recovery
4–23 kb: >80% recovery; 0.5–4 kb: >70% recovery;
50–500 bp: >50% recovery
>95% of linear DNA >22 bases
or bp; 99% retention of
unincorporated dye terminators
50–80%
Time required
<10 min
<10 min
<4 min
Freeze ’N Squeeze™ DNA Gel Extraction Spin Columns
see also
Agarose gel system:
pages 195–197.
DNA standards:
pages 144–147.
1
23
Freeze ’N Squeeze spin columns provide a quick and effective alternative
to chemical extraction and electroelution methods. The Freeze ’N Squeeze
method uses centrifugation to draw 50–23,000 bp DNA out of agarose gel
slices that have been quickly frozen and thawed. Features include:
• No
6
7
700
600
500
400
300
200
100
• DNA immediately available for PCR, subcloning, ligations, and
sequencing reactions
5
1,000 bp
solutions to prepare
• Less
4
Recovery of DNA fragments using Freeze ’N
Squeeze spin columns. Extracted fragments were
rerun on a ReadyAgarose™ 1% TAE gel. Lane 1,
precision molecular mass ruler; lane 7, EZ Load™
100 bp PCR molecular ruler.
than 1 minute of hands-on time
The Freeze ’N Squeeze Method
–20ºC for 5 min
Purified DNA
Cut out gel slice.
Chop into pieces.
Transfer to spin column
and freeze for 5 min.
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Spin for 3 min.
Recover purified DNA.
sample preparation
nucleic acid sample preparation
Ordering Information
Catalog #
732-6165
732-6166
Description
Price, US $
Freeze ’N Squeeze DNA Gel Extraction Spin Columns, 25
50.00
Freeze ’N Squeeze DNA Gel Extraction Spin Columns, 100145.00
Prepacked Spin Columns
Bio-Spin®, Micro Bio-Spin™, and PCR Kleen™ columns clean up DNA or protein samples quickly and easily. These
columns are available in multiple sizes and offer multiple MW exclusion limits to accommodate a variety of needs.
Use the chart below to choose the column that best meets your needs.
Prepacked Spin Column Selection Guide
Bio-Spin 6
Micro Bio-Spin 6
Bio-Spin 30
Micro Bio-Spin 30
PCR Kleen
Packed support
Special grade
Bio-Gel® P-6 gel
Special grade Bio-Gel P-6 gel
Special grade Bio-Gel P-30 gel
Special grade Bio-Gel P-30 gel
Special grade
size exclusion gel
Equilibration buffer10 mM Tris, pH 7.4 10 mM Tris, pH 7.4 10 mM Tris, pH 7.4 10 mM Tris, pH 7.410 mM Tris, 1 mM
or SSC buffer*
or SSC buffer*
or SSC buffer*
or SSC buffer*
EDTA, pH 7.0
Applications
Desalting of oligonucleotides
>20 bases
•
•
—
—
—
—
—
•
•
—
Removal of primers and primer-dimers from
PCR products >200 bp
—
—
—
—
•
•
•
—
•
—
DNA sequencing reaction mixture cleanup**
—
—
•
•
—
Riboprobe cleanup***
—
•
—
•
—
—
•
•
—
—
Purification of proteins of molecular weight
>6,000
•
•
—
—
—
Purification of proteins of molecular weight
>40,000
—
—
•
•
—
Labeling reactions: removal of unincorporated
nucleotides >20 bases
or bp from DNA
Buffer exchange (restriction fragments,
PCR products, enzyme
reactions, sequencing
templates)
Desalting of antibody, enzyme, and protein
solutions
Bed volume1.1 ml
0.7 ml1.1 ml
0.7 ml
0.6 ml
Retention and recovery
90% recovery of 20 bases or bp, 99% retention of salts
90% recovery of 20 bases or bp, 99% retention
of salts
95% recovery of 22 bases or bp, 98% retention
of ddNTPs
95% recovery of 22 bases or bp,
98% retention
of ddNTPs
85% recovery of
≥700 bp, 95%
retention of primers
and primer-dimers
Molecular weight exclusion limit,
globular proteins
6,000 6,000 40,000
40,000
8,000,000
Sample volume
50–100 µl10–75 µl
50–100 µl10–75 µl
25–100 µl
Centrifuge type
Swinging bucket Microcentrifuge
Swinging bucket Microcentrifuge
Microcentrifuge
Autoclavability
Yes
Yes
Yes
Yes
Yes
*150 mM NaCl, 17.5 mM sodium citrate, pH 7.0.
** In Tris buffer.
*** In RNase-free Tris buffer.
For more information, request bulletin 2320.
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sample preparation
nucleic acid sample preparation
Prepacked Bio-Spin and Micro Bio-Spin Columns
Bio-Spin and Micro Bio-Spin columns clean up DNA and protein samples quickly. Filled with specially
sized Bio-Gel P gels, Bio-Spin and Micro Bio-Spin columns are shipped fully hydrated in Tris or
SSC buffer and ready to use. These products effectively clean up and remove salts, nucleotides,
dye terminators, and small molecules from DNA, RNA, and protein samples. Features include:
• Convenience
of prepacked, prehydrated gel support
sample cleanup with a 4 minute microcentrifuge spin
• Efficient (98%) removal of ddNTPs and dye terminators
• Improved yield — 95% recovery of DNA >22 bp
• Sample loads from 10 to 100 µl
• Safe riboprobe preparation with RNase-free Micro Bio-Spin P-30 Tris spin columns
• Rapid
Empty columns are also available (pages 72–78). For more information on Bio-Gel P gels, see pages 57–58.
PCR Kleen Spin Columns
PCR Kleen columns are prepacked spin columns for purifying PCR products
and other DNA molecules >200 bp directly from reaction mixtures. A simple
4 minute spin effectively removes salts, nucleotides, enzymes, primers, and
primer-dimers. Purified DNA fragments are eluted into a collection tube
and are immediately available for secondary PCR, subcloning,
restriction digests, ligations, sequencing, and other
enzymatic manipulations.
For more information, request bulletin 2311.
Ordering Information
Catalog #
Description
Price, US $
Micro Bio-Spin Columns With Bio-Gel P-6 in Tris Buffer
732-6221
Micro Bio-Spin 6 Columns, includes 25 columns in Tris buffer, 50 collection tubes
70.00
Micro Bio-Spin 6 Columns, includes 100 columns in Tris buffer, 200 collection tubes 237.00
732-6222
Micro Bio-Spin 6 Columns, includes 1,000 columns in Tris buffer, 2,000 collection tubes 1,887.00
732-6225
Micro Bio-Spin Columns With Bio-Gel P-30 in Tris Buffer
Micro Bio-Spin 30 Columns, includes 25 columns in Tris buffer, 50 collection tubes
70.00
732-6223
Micro Bio-Spin 30 Columns, includes 100 columns in Tris buffer, 200 collection tubes
237.00
732-6224
Micro Bio-Spin 30 Columns, includes 1,000 columns in Tris buffer, 2,000 collection tubes1,887.00
732-6226
Micro Bio-Spin 30 Columns, includes 25 columns in Tris buffer, 50 collection tubes, RNase-free
83.00
732-6250
Micro Bio-Spin 30 Columns, includes 100 columns in Tris buffer, 200 collection tubes, RNase-free
282.00
732-6251
Micro Bio-Spin Columns With Bio-Gel P-6 in SSC Buffer
732-6200
Micro Bio-Spin 6 Columns, includes 25 columns in SSC buffer, 50 collection tubes
70.00
Micro Bio-Spin 6 Columns, includes 100 columns in SSC buffer, 200 collection tubes
237.00
732-6201
Micro Bio-Spin 6 Columns, includes 1,000 columns in SSC buffer, 2,000 collection tubes1,904.00
732-6205
Micro Bio-Spin Columns With Bio-Gel P-30 in SSC Buffer
732-6202
Micro Bio-Spin 30 Columns, includes 25 columns in SSC buffer, 50 collection tubes
70.00
Micro Bio-Spin 30 Columns, includes 100 columns in SSC buffer, 200 collection tubes
237.00
732-6203
Micro Bio-Spin 30 Columns, includes 1,000 columns in SSC buffer, 2,000 collection tubes1,904.00
732-6206
Bio-Spin Columns With Bio-Gel P-6 in Tris Buffer
Bio-Spin 6 Columns, includes 25 columns in Tris buffer, 50 collection tubes
86.00
732-6227
Bio-Spin 6 Columns, includes 100 columns in Tris buffer, 200 collection tubes
293.00
732-6228
Bio-Spin Columns With Bio-Gel P-30 in Tris Buffer
Bio-Spin 30 Columns, includes 25 columns in Tris buffer, 50 collection tubes
86.00
732-6231
Bio-Spin 30 Columns, includes 100 columns in Tris buffer, 200 collection tubes
293.00
732-6232
Bio-Spin Columns With Bio-Gel P-6 in SSC Buffer
Bio-Spin 6 Columns, includes 25 columns in SSC buffer, 50 collection tubes102.00
732-6002
Bio-Spin 6 Columns, includes 1,000 columns in SSC buffer, 2,000 collection tubes
2,445.00
732-6213
Bio-Spin Columns With Bio-Gel P-30 in SSC Buffer
732-6006
Bio-Spin 30 Columns, includes 25 columns in SSC buffer, 50 collection tubes102.00
PCR Kleen Spin Columns
732-6300
PCR Kleen Spin Columns, 25
10
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50.00
sample preparation
protein sample preparation
Protein Sample Preparation
Preparation of protein samples prior to downstream analysis is complex due to the variety of sample sources and strategies used to
clean or simplify them. Bio-Rad offers a suite of sample preparation products that provide solutions for a variety of needs.
• Chemical
reagent kits separate proteins based on differential solubilities of a sample’s components
techniques separate proteins based on their physical properties
• Electrophoretic techniques fractionate samples based on pI or MW
• Chromatographic
Bio-Rad sample preparation products can be used individually or together to cover any laboratory’s sample preparation needs, and
require minimum hands-on time. Use the diagram below to determine which sample preparation products best meet your needs.
For more information, go to www.bio-rad.com/proteinsampleprep/
* See pages 238–239.
Products for sample preparation.
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11
sample preparation
protein sample preparation
Protein Sample Preparation Kits
Bio-Rad offers sample preparation kits for a variety of needs. Protein enrichment kits (below) reduce high-abundance
proteins while enriching low-abundance proteins. Kits for cell lysis (page 13) lyse cells and extract proteins from a
variety of samples. General purpose kits (page 15) isolate total cellular protein, remove disulfide bonds, or remove salts
or other contaminants from proteins. Fractionation kits (page 16) reduce sample complexity by separating proteins by
cellular location, solubility, size, charge, and isoelectric point.
While these kits can be used to prepare samples for a variety of downstream protein analysis methods, many were
designed according to sample quantity and concentrations required for 1- and 2-D electrophoresis. Stringent quality
control and preparation of samples prior to 2-D electrophoresis greatly improves reproducibility and resolution.
Protein Enrichment
see also
Aurum serum protein
mini kit: page 15.
Biomarker Research
Center services:
page 310.
ProteoMiner™ Protein Enrichment Kits new
High-abundance proteins
present in complex biological
samples such as sera or
plasma make the detection of
medium- and low-abundance
proteins extremely challenging.
ProteoMiner protein enrichment
technology is a novel sample
preparation tool used for the
compression of the dynamic
range of proteomes from
complex biological samples,
allowing the exploration of the
entire proteome.
ProteoMiner protein enrichment technology employs a
large, highly diverse bead-based library of combinatorial
peptide ligands, which simultaneously dilutes highabundance proteins and concentrates low-abundance
proteins. When a complex biological sample is applied to
the beads, high-abundance proteins saturate their highaffinity ligand(s), and excess protein is washed away.
In contrast, low-abundance proteins are concentrated
on their specific affinity ligand(s).
Benefits of ProteoMiner protein enrichment kits:
• Decreases
the amount of high-abundance proteins
without immunodepletion — prevents the loss of
proteins bound to high-abundance proteins
• Enriches
medium- and low-abundance proteins that
cannot be detected through traditional methods
12
ProteoMiner technology. Each bead features a different hexapeptide
ligand with affinity for specific proteins in a sample. Samples are applied
to the beads, allowing proteins to bind to their specific ligands. Proteins
in excess are washed away, and those proteins bound to the beads are
eventually eluted, allowing further downstream analysis.
• Compatible
with a variety of sample types
• Compatible
with all major downstream protein analysis techniques
• Convenient,
easy-to-use format
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sample preparation
protein sample preparation
ProteoMiner Protein Enrichment Kit
The ProteoMiner protein enrichment kit provides all
necessary reagents for accessing low-abundance
proteins, utilizing a single elution reagent in a simple,
easy-to-perform process. This kit can be used with a
variety of biological samples and is compatible with all
major downstream proteomics applications.
Untreated plasma
Treated plasma
ProteoMiner Sequential Elution Kit
The ProteoMiner sequential elution kit utilizes multiple
elution reagents to sequentially elute proteins based
on different properties. This kit fractionates proteins
to improve detection and resolution and has been
optimized for SELDI experiments.
Untreated serum
Treated serum
For more information, go to
www.bio-rad.com/proteominer/
Reduction of high-abundance proteins improves detection and
resolution of proteins. Left, untreated plasma and serum; right, plasma
and serum treated using the ProteoMiner protein enrichment kit.
Ordering Information
Catalog #
Description
163-3000
ProteoMiner Protein Enrichment Kit, 10 preps, includes 10 spin columns, wash buffer, elution reagents, 650.00
collection tubes
ProteoMiner Introductory Kit, 2 preps, includes 2 spin columns, wash buffer, elution reagents, collection tubes175.00
ProteoMiner Sequential Elution Kit, 10 preps, includes 10 spin columns, wash buffer, 4 sequential elution 750.00
reagents, collection tubes
ProteoMiner Sequential Elution Reagents, includes reagents only (columns not included), to be used 160.00
with 163-3000
163-3001
163-3002
163-3003
Price, US $
Kits for Cell Lysis
see also
MicroRotofor™ Lysis Kits
Reducing protein sample complexity allows
visualization of low-abundance proteins. An effective
method for reducing sample complexity is through
protein sample fractionation using IEF.
MicroRotofor cell:
page 236.
For easy small-scale IEF, MicroRotofor cell lysis
and protein extraction kits are available to prepare
samples for the MicroRotofor cell, which performs
liquid-phase IEF with only 2.5 ml. The kits are
convenient and easy to use, and provide tailored
protocols based on sample type. All kits are quality
controlled to guarantee performance and help
ensure reproducibility of IEF runs.
• Cell
lysis and extraction protocols are tailored for mammalian, plant, yeast, and bacterial samples
•Kits
can be used for any species-specific protein extraction — a MicroRotofor run is not a necessary downstream
application
•Resulting
protein extract can be run immediately in the MicroRotofor cell — no need to change buffer conditions to
accommodate the run
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13
sample preparation
protein sample preparation
ReadyPrep™ Mini Grinders
ReadyPrep mini grinders allow grinding of small biological samples for
high recovery of proteins and nucleic acids. Each mini
grinder includes a 1.5 ml grinding tube containing a
grinding resin and a matching pestle.
The grinding resin is a neutral abrasive
material made of high tensile-strength
microparticles that do not bind proteins
or nucleic acids. ReadyPrep mini grinders
are disposable and nuclease- and
protease-free. They are a component of
the MicroRotofor lysis kit (mammal) and are
also sold separately as a pack of 20. The mini grinders:
• Are
convenient for small-scale extraction from biological tissues
an inert grinding resin that facilitates efficient tissue disruption
• Fit conveniently in most benchtop centrifuges
•Contain
Bio-Plex® Cell Lysis Kit
The Bio-Plex cell lysis kit has been developed specifically
to prepare cell culture and tissue lysate samples for analysis
with Bio-Plex phosphoprotein and total target assays. This
cell lysis kit can also be used to prepare cell lysates for western
blot analysis. Its protein extraction procedure yields western
blotting results similar to those generated by routine cell lysis and
protein extraction protocols.
Ordering Information
Catalog #
Description
MicroRotofor Lysis Kits
163-2141 MicroRotofor Cell Lysis Kit (Mammal), 15 preps, includes 50 ml protein solubilization buffer (PSB), ReadyPrep mini grinders (2 packs of 10 each)
MicroRotofor Cell Lysis Kit (Plant), 10 preps, includes 50 ml protein solubilization buffer (PSB), 163-2142 ReadyPrep 2-D cleanup kit (50 reaction size)
MicroRotofor Cell Lysis Kit (Yeast), 15 preps, includes 50 ml protein solubilization buffer (PSB), 163-2143 15 ml yeast suspension buffer, 2 x 0.5 ml lyticase (1.5 U/µl)
MicroRotofor Cell Lysis Kit (Bacteria), 15 preps, includes 50 ml protein solubilization buffer (PSB), 163-2144 25 ml bacteria suspension buffer, 1 ml lysozyme (1,500 U/µl)
MicroRotofor Lysis Kit Components
ReadyPrep Mini Grinders, includes 2 sealed bags of 10 mini grinders each, sufficient for twenty 100 mg extractions
163-2146 Protein Solubilization Buffer (PSB), makes 50 ml of solution
163-2145 Price, US $
210.00
270.00
230.00
230.00
95.00
95.00
Bio-Plex Cell Lysis Kit
171-304011 Bio-Plex Cell Lysis Kit, 1 x 96-well154.00
Bio-Plex Cell Lysis Kit, 10 x 96-well1,287.00
171-304012 14
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sample preparation
protein sample preparation
General Purpose Cleanup Kits
General purpose cleanup kits are available for the removal of salts, high-abundance proteins, and other contaminants. These kits
improve sample quality, thus improving results in downstream applications.
General Purpose Sample Preparation Kit Selection Guide
Kit Type and Catalog Number
Applications
Procedure
Preparation
Acceptable
Number of
Time
Sample Sources
Preps
Salt Removal
ReadyPrep™ 2-D cleanup kit 163-2130
Reduction of streaking on 2-D gels
Concentration of dilute samples
TCA-like precipitation to remove salts, <1 hr
s s s s
detergents, lipids, phenolic compounds
50
Micro Bio-Spin™ P-6 columns
Removal of salts and other contaminants Size exclusion chromatography
5 min —
25, 100,
(see page 9)1,000
Albumin/IgG Reduction/Removal
ProteoMiner™ protein enrichment kit Reduction of high-abundance proteins and Affinity chromatography3 hr
(see page 12)
enrichment of low-abundance proteins
Aurum™ serum protein mini kit
732-6713, 732-6701 s s s s
Simultaneous removal of albumin Affinity chromatography
<30 min s
and IgG
Aurum™ Affi-Gel® Blue mini columns Removal of up to 90% albumin
732-6708, 732-6709
Affinity chromatography
Reduction of streaking on 2-D gels
TCA-like precipitation to remove <1 hr
by removal of detergents, phenolic
salts, detergents, lipids, phenolic
compounds, salts, and most other ionic compounds
contaminants
Concentration of dilute samples
ReadyPrep reduction-alkylation kit Reduction of streaking on 2-D gels
163-2090 Improved resolution of basic proteins
ReadyPrep protein extraction kit
Isolation of total cellular protein
(total protein) 163-2086 2, 10
<30 min s
Removal of Other Contaminants
ReadyPrep 2-D cleanup kits 163-2130, 163-2140 2, 10
2, bulk
s s s s
50
Reduction, then alkylation of sample
<2 hr
to remove disulfide bonds and prevent
their re-formation
s s s s100
Total protein solubilization using
strong detergent ASB-14
s s* s s
<1 hr
20
s Mammalian tissue or cells. s Plant leaves. s E. coli. s Yeast. s Serum, plasma.
* Plant leaves can be a sample source, but the 2-D cleanup kit must be used after the total protein kit.
Better 2-D resolution and reproducibility — Improving
resolution and ensuring reproducibility are two
important goals for any 2-D experiment, and protein
sample preparation plays a key role in achieving
those goals. ReadyPrep and Aurum kits provide tools
and proven protocols to improve 2-D gel resolution
by reducing smearing or sample complexity. The
ReadyPrep kits prepare protein samples using solution
chemistry and differential solubility, while the Aurum
kits are column-based. Regardless of which kit you
choose, stringent quality control ensures consistently
high-quality products for reproducible results. Refer
to the guides on the previous page for an overview of
these kits.
For more information on ReadyPrep sample preparation
kits, request bulletins 2934, 2961, and 3006. For
more information on the Aurum serum protein mini kit,
request bulletin 2823.
Before
After
Effective removal of detergents using the ReadyPrep 2-D cleanup kit leads to improved
2-D SDS-PAGE separation. E. coli extracts were spiked with 1% SDS. Left, untreated
sample; right, sample treated with the ReadyPrep kit.
Before
After
Albumin
Heavychain
IgG
Lightchain
IgG
Albumin
Heavychain
IgG
Lightchain
IgG
Removal of albumin and IgG from serum using the Aurum serum protein mini kit.
Total protein (1.32 mg) was purified on an Aurum serum protein mini column. Left, untreated
serum; right, serum treated with the Aurum kit.
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15
sample preparation
protein sample preparation
Ordering Information
Catalog #
Description
Price, US $
General Purpose Kits
163-2130
ReadyPrep 2-D Cleanup Kit, 50 preps 213.00
ReadyPrep 2-D Starter Kit, includes protein sample and reagents sufficient to rehydrate, focus, and transfer to second-dimension 151.00
163-2105
gels or six 17 cm, ten 11 cm, or sixteen 7 cm ReadyStrip IPG strips (ReadyStrip IPG strips and precast SDS-PAGE gels and
gel stains not included in kit)
ReadyPrep Reduction-Alkylation Kit, 100 preps134.00
163-2090
ReadyPrep Protein Extraction Kit (Total Protein), 20 preps122.00
163-2086
Aurum Serum Protein Mini Kit, 2 pack, includes columns and buffers
72.00
732-6713
Aurum Serum Protein Mini Kit, 10 pack, includes columns and buffers337.00
732-6701
Aurum Affi-Gel Blue Mini Kit, 2 pack, includes columns and buffers
56.00
732-6712
Aurum Affi-Gel Blue Mini Columns, 25 pack
248.00
732-6708
Aurum Affi-Gel Blue Mini Columns, 100 pack
442.00
732-6709
ReadyPrep Kit Components and Related Products
163-2091
ReadyPrep Proteomics Grade Water, 500 ml31.00
ReadyPrep Sequential Extraction Kit Reagent 1, 50 ml, 40 mM Tris base
23.00
163-2102
ReadyPrep Sequential Extraction Kit Reagent 2, 10 ml, 8 M urea, 4% CHAPS, 40 mM Tris, 0.2% Bio-Lyte 3/10 ampholyte
28.00
163-2103
ReadyPrep Sequential Extraction Kit Reagent 3, 10 ml, 5 M urea, 2 M thiourea, 2% CHAPS, 2% SB 3-10, 40 mM Tris, 28.00
163-2104
0.2% Bio-Lyte 3/10 ampholyte
ReadyPrep 2-D Rehydration/Sample Buffer 1, 10 ml, 7 M urea, 2 M thiourea, 1% ASB-14, 40 mM Tris
27.00
163-2083
ReadyPrep 2-D Starter Kit Rehydration/Sample Buffer, 10 ml, 8 M urea, 2% CHAPS, 50 mM DTT, 23.00
163-2106
0.2% Bio-Lyte 3/10 ampholyte, 0.001% Bromophenol Blue
Reducing and Alkylating Agents
Dithiothreitol (DTT), 5 g
64.00
161-0611
Tributylphosphine (TBP), 200 mM, 0.6 ml33.00
163-2101
Iodoacetamide, 30 g102.00
163-2109
Fractionation Kits
Fractionation kits reduce sample complexity, helping to identify low-abundance proteins. Fractionation kits can be subdivided into
three groups, which fractionate based on: cellular location of the proteins of interest, differential solubility, and protein charge.
Fractionation Sample Preparation Kit Selection Guide
Kit Type and Catalog Number
Applications
Procedure
Differential Solubility
ReadyPrep™ sequential extraction kit
163-2100 2-D, 1-D SDS-PAGE
Differential solubilization with SB 3-10
<3 hr
(Molloy et al. 1998)
s s s15
Division of whole proteome into
3 fractions
Differential solubilization with strong
<1 hr
detergent ASB-14
s s s 20
Organelle isolation followed by
<3 hr
s s
strongly chaotropic extraction buffer
(Dignam et al. 1983, Zerivitz and
Akusjarvi 1989)
50
Temperature-dependent differential
<3 hr
solubilization with Triton X-114
(Bordier 1981)
50
ReadyPrep protein extraction
kit (soluble/insoluble)
163-2085 2-D, 1-D SDS-PAGE
Division of whole proteome into
2 fractions
Cellular Location
ReadyPrep protein extraction
2-D, 1-D SDS-PAGE, western blotting
kit (cytoplasmic/nuclear)
Isolation of cytoplasmic and nuclear 163-2089
proteins from eukaryotic samples
ReadyPrep protein extraction
2-D, 1-D SDS-PAGE, western blotting
kit (membrane I)
Isolation of membrane proteins
163-2088 Preparation
Acceptable
Number of
Time
Sample Sources
Preps
s s s s ReadyPrep protein extraction
2-D, 1-D SDS-PAGE, western blotting Sodium carbonate extraction3 hr
and ultracentrifugation (Fujiki et al. kit (membrane II)
Isolation of membrane proteins
163-2084 1982, Molloy et al. 2000)
s s s10
ReadyPrep protein extraction
kit (signal) 163-2087 s s s
Protein Charge
Aurum™ AEX and CEX mini kits
(see page 18)
2-D, 1-D SDS-PAGE, western blotting
Differential solubilization with
<3 hr
Isolation of signal (caveolin-associated) Triton X-100 (Brown and Rose 1992,
Simons and Ikonen 1997)
proteins
Enrich for either acidic or basic
Ion exchange chromatography
<30 min
s s s ss
proteins
s Mammalian tissue or cells. s Plant leaves. s E. coli. s Yeast. s Serum, plasma.
16
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50
2, bulk
sample preparation
protein sample preparation
Three kits to choose from for membrane proteins — Bio-Rad offers three ReadyPrep protein extraction kits to
facilitate the isolation of membrane proteins. The ReadyPrep protein extraction kits (membrane I and II) yield fractions
of varying complexity. The membrane I kit is a quick and effective protocol for isolating most membrane proteins.
It does not require ultracentrifugation or preparation of density gradients. The membrane II protocol is better for
isolating more complex membrane proteins. The ReadyPrep protein extraction kit (signal) is specific for isolating
proteins involved in intracellular membrane trafficking and signaling pathways. These include proteins such as
GPI-anchored proteins, caveolin and associated proteins, acylated tyrosine kinases, and G proteins.
1-D family
Optimizing every
step of the 1-D
electrophoresis and
blotting workflow:
page 136.
Fractionation kits using differential protein solubility — The ReadyPrep sequential extraction kit and the
ReadyPrep protein extraction kit (soluble/insoluble) both reduce sample complexity using differential solubilization.
The sequential extraction kit enables the isolation of three different fractions of increasing solubility. These fractions
are isolated sequentially, allowing the visualization of proteins that might not otherwise be seen. Increasing
solubilization strength is provided through the use of stronger detergents for each subsequent fraction. In the
ReadyPrep sequential extraction kit, the solution with the strongest solubilizing power is reagent 3, which contains
the detergent SB 3-10. In contrast, the ReadyPrep protein extraction kit (soluble/insoluble) uses a single fractionation
step and uses the detergent ASB-14, a stronger detergent than SB 3-10. The two kits can be used independently,
or the rehydration/sample buffer from the soluble/insoluble kit can be used with the sequential extraction kit to create
a fourth fraction for even better resolution.
Step 1
Step 2
Step 3
The ReadyPrep sequential extraction kit. The kit distributes proteins into three different fractions based on differential solubility, providing increased
resolution of proteins on 2-D gels.
Fractionation by charge — Chromatographic methods also can fractionate specifically or generally. Aurum ion
exchange (AEX and CEX) mini columns allow selective purification of either acidic or basic proteins. Easy-to-use Aurum
ion exchange kits selectively enrich either acidic or basic proteins and can be used with a variety of starting samples.
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17
sample preparation
protein sample preparation
Total protein sample
AEX-bound sample
CEX-bound sample
pI
3 10
pI
3 10
pI
3 10
Fractionation of rat brain tissue using Aurum ion exchange mini columns. Rat brain total protein extracts (3 ml) were loaded onto an Aurum AEX
or CEX mini column, then eluted using the recommended protocol. The fractionated samples were treated with the ReadyPrep reduction/alkylation
and 2-D cleanup kits prior to 2-D gel analysis. Left, total rat brain protein; center, sample that bound to the Aurum AEX column (acidic proteins); right,
sample that bound to the Aurum CEX column (basic proteins).
For more information on ReadyPrep kits, request bulletins 2934, 2961, and 3006. For more information on Aurum ion
exchange kits, request bulletin 2928.
ReadyPrep Kit Components and Related Products
Individual ReadyPrep kit components and related products such as reducing agents are also available.
See ordering information.
Ordering Information
Catalog #
Description
Price, US $
Fractionation Kits
163-2100
ReadyPrep Sequential Extraction Kit, 15 preps151.00
ReadyPrep Protein Extraction Kit (Soluble/Insoluble), 20 preps160.00
163-2085
ReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear), 50 preps308.00
163-2089
ReadyPrep Protein Extraction Kit (Membrane I), 50 preps308.00
163-2088
ReadyPrep Protein Extraction Kit (Membrane II), 10 preps183.00
163-2084
ReadyPrep Protein Extraction Kit (Signal), 50 preps156.00
163-2087
Aurum CEX Mini Kit, 2 pack, includes columns and buffers
56.00
732-6711
Aurum CEX Mini Columns, 25 pack
248.00
732-6703
Aurum CEX Mini Columns, 100 pack
442.00
732-6704
Aurum AEX Mini Kit, 2 pack, includes columns and buffers
56.00
732-6710
Aurum AEX Mini Columns, 25 pack
248.00
732-6706
Aurum AEX Mini Columns, 100 pack
442.00
732-6707
ReadyPrep Kit Components and Related Products
163-2102
ReadyPrep Sequential Extraction Kit Reagent 1, 50 ml, 40 mM Tris base
ReadyPrep Sequential Extraction Kit Reagent 2, 10 ml, 8 M urea, 4% CHAPS, 40 mM Tris, 163-2103
0.2% Bio-Lyte 3/10 ampholyte
ReadyPrep Sequential Extraction Kit Reagent 3, 10 ml, 5 M urea, 2 M thiourea, 2% CHAPS, 2% SB 3-10, 163-2104
40 mM Tris, 0.2% Bio-Lyte 3/10 ampholyte
18
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23.00
28.00
28.00
sample preparation
protein sample preparation
Protein Sample Preparation Instruments
Bio-Rad offers apparatus for prefractionation of samples, resulting in increased resolution of proteins, enrichment
of low-abundance proteins, and the ultimate identification of more proteins on a 2-D gel. You have the option of
purification by chromatography or preparative electrophoresis.
Bio-Rad offers a complete line of chromatography instruments for high-resolution purification. Alternatively,
preparative electrophoresis devices can be combined with other separation techniques to provide sample
preparation schemes for numerous proteomic applications. Use preparative electrophoresis products, such as
the Model 491 and mini prep cells or the Rotofor® family of cells (see below), to purify and fractionate up to gram
quantities of proteins by molecular weight, pI, or a combination of both.
see also
RC DC protein assay
kits: pages 31–32.
Chromatography Systems
A complete line of chromatography instruments meets Bio-Rad’s continuing commitment to offer high-quality
products that are flexible, upgradable, and easy to use. These biocompatible systems include low-pressure
Econo™ components, the low-pressure BioLogic™ LP system, and the high-resolution BioLogic DuoFlow™ systems.
For more information, see pages 100–121.
Model 491 Prep Cell and Mini Prep Cell
The Model 491 prep cell and mini prep cell use continuous-elution electrophoresis to purify macromolecules based
on their relative mobility in polyacrylamide or agarose gels. Both the Model 491 prep cell and the mini prep cell allow
resolution of proteins differing in molecular weight by as little as 2%. DNA fragments up to 18 kb may also be purified
in a prep cell using agarose gel electrophoresis.
For more information, see pages 238–239.
MicroRotofor™ Cell, Mini Rotofor® Cell, and Rotofor Cell
The Rotofor family of products fractionates complex protein samples that are free in solution. The nondenaturing
liquid-phase IEF technique rapidly isolates proteins of interest based on pI, simplifying protein purification and
maintaining biological activity. The MicroRotofor, mini Rotofor, and Rotofor cells integrate into any downstream
workflow, and are ideal primary purification tools.
For more information, see pages 236–238.
Protein Sample Preparation
Cartridges and Columns
Bio-Rad’s prepacked chromatography columns and cartridges offer a quick and convenient way to experiment
with different separation techniques when the protein to be purified has not yet been characterized. The prepacked
cartridges are for use with the BioLogic™ systems, a peristaltic pump, or any chromatography system. For simple
step elution, the cartridges can be used with a syringe.
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see also
Bottled media:
pages 36–61.
BioLogic systems:
pages 101–114.
Econo pump:
page 116.
19
sample preparation
protein sample preparation
Bio-Scale™ Mini Cartridges
Bio-Scale Mini cartridges have a double-wall cartridge
design* that provides extra durability and allows easy,
reliable runs with aqueous buffers most commonly used for
protein separation, at pressures to 45 psi. The cartridge
selection includes UNOsphere™ Q, UNOsphere S, and
Profinity™ IMAC media prepacked into 1 and 5 ml formats.
Bio-Scale Mini cartridges are convenient, disposable, and
supplied ready for use.
The cartridges are designed for use with BioLogic™ systems,
a peristaltic pump, or any chromatography system. Bio-Scale Mini
affinity cartridges can also be run on the Profinia™ protein
purification system (see pages 86–93). For simple step elution,
the cartridges can be used with a Luer-Lok syringe. Fittings are
available for connection to HPLC- and FPLC-type systems.
Other features include:
•Simple
luer fittings for easy connection to any system
compatible polypropylene parts
• A wide range of chromatography media
• Chemically
For more information, see pages 63–64 or visit www.bio-rad.com/cartridges/
Bio-Spin® and Micro Bio-Spin™ Columns
Bio-Spin and Micro Bio-Spin columns are fully hydrated and ready to use for the quick
cleanup of protein samples and DNA samples.
3.75 cm
For a list of applications and prefilled column ordering information, see page 65.
Empty columns are also available; see page 72.
6 cm
Poly-Prep Columns
®
These convenient 2 µl gravity-flow chromatography columns are useful
for sample preparation and small-scale chromatography applications,
including work with radioisotopes and other applications that require
disposable products.
9 cm
For more information, see page 65 for prepacked columns and page 73 for empty columns.
Econo-Pac® Columns
Econo-Pac columns are used for gravity-flow chromatography.
For open column work, a special upper bed support prevents
the bed from running dry. Bed volumes from 1–20 ml are
acceptable. These columns can be autoclaved and will retain
fine particles of 30 µm and larger.
14 cm
For more information, see page 66 for prepacked columns and pages 72–73 for empty columns.
* Patent pending.
20
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