Product Datasheet Anti-CD11b antibody [M1/70] ab8878 10 Abreviews 18 References 3 Images Overview Product name Anti-CD11b antibody [M1/70] Description Rat monoclonal [M1/70] to CD11b Tested applications ICC/IF, Flow Cyt Species reactivity Reacts with: Mouse, Rabbit, Human Immunogen The hybridoma was formed by the fusion of mouse myeloma NS1 cellswith spleen cells from rats immunized with B10 mouse spleen cells enrichedfor T lymphocytes. General notes CD11b is also known as MAC-1alpha, CR3 and integrin alpha M. We do not batch test ab8878 in IHC, however, some customers have successfully used this antibody in this application. We cannot recommend this antibody for IHC. Properties Form Liquid Storage instructions Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. Storage buffer pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Purity Protein G purified Clonality Monoclonal Clone number M1/70 Myeloma NS1 Isotype IgG2b Light chain type unknown Applications Our Abpromise guarantee covers the use of ab8878 in the following tested applications. The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user. Application Abreviews Notes ICC/IF Use a concentration of 5 µg/ml. Flow Cyt Use at an assay dependent concentration. Each labeling step was carried out for 30 min at 0-4°C in mediumcontaining 0.01 M NaN3. Cells (5 x 107/ml) were incubated with 50 µl M1 monoclonal antibody or irrelevant monoclonal antibody, R4/18.2, as control in the first step, washed, suspended in 50 µl of FITC-F (ab')2 anti-rat IgGin the second step, and washed through a layer of fetal calf serum. Can also be used in cytotoxicity and binding assays. Target Function Integrin alpha-M/beta-2 is implicated in various adhesive interactions of monocytes, macrophages and granulocytes as well as in mediating the uptake of complement-coated particles. It is identical with CR-3, the receptor for the iC3b fragment of the third complement component. It probably recognizes the R-G-D peptide in C3b. Integrin alpha-M/beta-2 is also a receptor for fibrinogen, factor X and ICAM1. It recognizes P1 and P2 peptides of fibrinogen gamma chain. Tissue specificity Predominantly expressed in monocytes and granulocytes. Involvement in disease Genetic variations in ITGAM has been associated with susceptibility to systemic lupus erythematosus type 6 http://www.abcam.com/CD11b-antibody-M170-ab8878.html 1 Product Datasheet (SLEB6) [MIM:609939]. Systemic lupus erythematosus (SLE) is a chronic, inflammatory and often febrile multisystemic disorder of connective tissue. It affects principally the skin, joints, kidneys and serosal membranes. It is thought to represent a failure of the regulatory mechanisms of the autoimmune system. Sequence similarities Belongs to the integrin alpha chain family. Contains 7 FG-GAP repeats. Contains 1 VWFA domain. Domain The integrin I-domain (insert) is a VWFA domain. Integrins with I-domains do not undergo protease cleavage. Cellular localization Membrane. Anti-CD11b antibody [M1/70] images ICC/IF image of ab8878 stained Raw 246.7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA/ 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab8878 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rat IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGAwas used Immunocytochemistry/ Immunofluorescence - AntiCD11b antibody [M1/70] (ab8878) to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. Overlay histogram showing Human macrophages stained with ab8878 (blue line). Cells were incubated with human immunoglobulin for 30 min at 4°C to block nonspecific protein-protein interactions. The cells were then incubated with the antibody (ab8878, 20 µg/mL) for 30 min at 4ºC. The secondary antibody used was a mouse anti-rat FITC (IgG2b gamma chain) (ab99671) at 1 µg/mL for 30 min at 4ºC. Isotype control antibody (black line) was Rat IgG2b [RTK4530] (ab18541) used under the same conditions. Flow Cytometry - Anti-CD11b [M1/70] antibody (ab8878) http://www.abcam.com/CD11b-antibody-M170-ab8878.html 2 Product Datasheet Overlay histogram showing Human macrophages stained with ab8878 (red line). Cells were pre-incubated in Human AB serum (10%) for 20 mins at 4ºC. Cells were then incubated with the antibody (ab8878, 0.1µg/1x10^6 cells) for 30 min at 4ºC. The secondary antibody used was a goat anti-rat Alexa Fluor® 488 (IgG H+L) (ab150165) at 1/2000 dilution for 30 min at 4ºC. Isotype control antibody (black line) was rat IgG2b [RTK4530] (ab18541, 1µg/1x10^6 cells) used under the same conditions. Unlabelled sample (blue line) was also Flow Cytometry - Anti-CD11b [M1/70] antibody used as a control. Acquisition of >5,000 events were (ab8878) collected using a 20mWArgon ion laser (488nm) and 525/30 bandpass filter. 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