BD Horizon™ V500-C Antibody Conjugates Violet Laser Reagents Features ptimized for compensation setup using O BD™ Multicolor CompBeads educes spillover into the FITC channel compared to R AmCyan aximizes choice and flexibility for multicolor panel M design Available as CE-IVD and RUO(GMP) reagents 200 250 S upports 8-color analysis on the BD FACSCanto™ II flow cytometer (x 1,000) BD Biosciences continues to expand the options for multicolor flow panel design with the development of an optimized BD Horizon™ brand dye for the violet laser. The BD Horizon V500-C dye has been specifically engineered to reduce spectral overlap into the FITC channel (Table 1) and to be compatible with BD™ Multicolor CompBeads particles for establishing compensation values (Table 2). BD Horizon V500-C is an organic dye that offers the same stability and compatibility typical of the BD Horizon family of dyes. Optimized for the violet laser BD Horizon V500-C is a completely novel organic dye structure that has been specifically engineered to have optimized spectral characteristics for use with the violet laser. It is optimally suited for use with BD FACS™ brand flow cytometers equipped with violet lasers such as the BD FACSCanto II flow cytometer. When used with these instruments, BD Horizon V500-C reagents demonstrate improved spectral characteristics compared to other violet laser excitable dyes, and enables advanced multicolor panels together with the BD Horizon V450 reagents. 50 100 SSC 150 With a maximum excitation at 415 nm and an emission peak at 500 nm, BD Horizon V500-C is readily compatible with the standard filter sets on BD FACS brand flow cytometers equipped with violet lasers. The addition of BD Horizon V500-C increases fluorochrome choice by enabling 8-color analysis on the BD FACSCanto II flow cytometer. Lymphocytes 0101 102 103 104 105 105 CD45 V500-C Expand your multicolor capability Designed with your current protocols in mind, BD Horizon V500-C reagents demonstrate compatibility with the standard staining buffers typically used in flow cytometry. Compatibility with red cell lysis buffers such as BD FACS™ lysing solution as well as permeabilization buffers for intracellular staining (eg, BD FACS™ Permeabilizing Solution 2 and BD Intrasure™) has been shown. BD Horizon V500-C shows stability in the presence of paraformaldehyde-based fixatives and its 5 μL/test formulation simplifies creation of multicolor cocktails. 0 102 103 CD4 FITC 104 Visit bdbiosciences.com for more information. -102 0 102 103 104 105 CD3 APC-H7 Figure 1. Lysed whole blood gated on CD45 lymphocytes and subsequent resolution of T cell subsets. BD Horizon™ V500-C Antibody Conjugates Violet Laser Reagents Instrument Excitation Laser Line (nm) Fluorescence Channel BD FACSCanto II 488 633 405 Fluorochromes provided by BD Biosciences Green Yellow FITC PE Red PerCP Infrared PE-Cy™7 Red APC Infrared APC-H7 APC-Cy7* Green Blue BD Horizon V500-C BD Horizon V450 BD Horizon V500 PerCP-Cy™5.5 Figure 2. BD Biosciences Fluorochrome Reference Chart To select the optimal combination of fluorochromes, visit bdbiosciences.com/spectra to use the interactive fluorescence spectrum tool. V500-C SOVs into… FITC Beads % Spillover into FITC Description V500-C AmCyan CD3 8.6% 43.4% CD45 8.0% 44.7% Table 1. BD Horizon V500-C conjugates have significantly less spillover into the FITC channel compared to AmCyan. Comparison of lysed whole blood stained with anti-CD3 or anti-CD45 to establish SOV on a BD FACSCanto™ II flow cytometer. V450 Absolute Cells Difference Beads (Beads - Cells) Cells Absolute Difference (Beads - Cells) Average 8.1 8.3 0.4 10.7 11.9 1.2 Average Max-Min/ Reagent 0.5 0.2 0.3 0.6 0.6 0.3 Total Max-Min All Lots 1.1 1.2 0.6 1.2 1.2 1.0 Table 2. SOVs for V500-C can be obtained with BD CompBeads or cells. The average is the value across 18 independent lots of V500-C reagents (three lots each of six different reagents) measured using BD CompBeads or cellular controls. The average Max-Min/Reagent illustrates the average variation (difference in the maximum and minimum SOV) obtained within the three lots for each reagent. The total Max-Min (all lots) represents the total variation (difference in the maximum and minimum SOV) obtained across all 18 lots. BD Horizon™ V500-C CE-IVD(1) Reagents Description React. Clone Isotype Size Reg. Status Cat. No. CD3 Hu SK7 Ms IgG1 100 tests CE-IVD 652896 CD45 Hu 2D1 Ms IgG1 100 tests CE-IVD 655873 Ordering Information BD Horizon™ V500-C RUO(GMP)(2) Reagents Description React. Clone Isotype Size Reg. Status Cat. No. CD3 Hu SK7 Ms IgG1, k 100 tests RUO (GMP) 647453 CD4 Hu SK3 Ms IgG1, k 100 tests RUO (GMP) 647455 CD8 Hu SK1 Ms IgG1, k 100 tests RUO (GMP) 647457 CD14 Hu MfP9 Ms IgG2b, k 100 tests RUO (GMP) 647459 CD20 Hu L27 Ms IgG1, k 100 tests RUO (GMP) 647463 CD45 Hu 2D1 Ms IgG1, k 100 tests RUO (GMP) 647449 CD138 Hu MI15 Ms IgG1, k 100 tests RUO (GMP) 650659 X40 Ms IgG1, k 100 tests RUO (GMP) 647451 Mouse IgG1 CE-IVD: For in vitro diagnostic use. CE marked in conformity with the European IVD Directive 98/79/EC (Directive on in vitro diagnostic medical devices). (1) RUO(GMP): For Research Use Only (General Manufacturing Practices). Not for use in diagnostic or therapeutic procedures. (2) *APC-Cy7: US Patent 5,714,386 Cy™ is a trademark of Amersham Biosciences Corp. Cy™ dyes are subject to proprietary rights of Amersham Biosciences Corp and Carnegie Mellon University and are made and sold under license from Amersham Biosciences Corp only for research and in vitro diagnostic use. Any other use requires a commercial sublicense from Amersham Biosciences Corp, 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA. BD, BD Logo and all other trademarks are property of Becton, Dickinson and Company. © 2012 BD XEUR0296-00 BD Biosciences bdbiosciences.com/eu
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