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PerCP-Cy™5.5 Mouse Anti-Human HLA-DR
PerCP-Cy™5.5 Mouse Anti-Human HLA-DR

Flow cytometric analysis of HLA-DR expression on Rhesus macaque (Macaca mulatta) peripheral blood leucocytes.  Rhesus whole blood was stained with either PerCP-Cy™5.5 Mouse IgG2a, κ Isotype Control (Cat. No. 552577; dashed line histogram) or PerCP-Cy™5.5 Mouse Anti-Human HLA-DR (Cat. No. 560652/552764; solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Fluorescent histograms showing expression of HLA-DR (or Ig Isotype control staining) were derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations.

Flow cytometric analysis of HLA-DR expression on Rhesus macaque (Macaca mulatta) peripheral blood leucocytes.  Rhesus whole blood was stained with either PerCP-Cy™5.5 Mouse IgG2a, κ Isotype Control (Cat. No. 552577; dashed line histogram) or PerCP-Cy™5.5 Mouse Anti-Human HLA-DR (Cat. No. 560652/552764; solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Fluorescent histograms showing expression of HLA-DR (or Ig Isotype control staining) were derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations.

Product Details
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BD Pharmingen™
MHC class II antigen; HLA class II histocompatibility antigen
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development), Dog (Reported)
Mouse IgG2a, κ
Flow cytometry (Routinely Tested)
20 µl
AB_394453
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  7. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  8. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  9. Cy is a trademark of GE Healthcare.
  10. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
552764 Rev. 8
Antibody Details
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G46-6

The G46-6 monoclonal antibody specifically binds to HLA-DR, a major histocompatibility complex (MHC) class II antigen. HLA-DR antigens are encoded by genes within the Human Leukocyte Antigen (HLA) Complex located on chromosome 6. HLA-DR is a transmembrane heterodimeric glycoprotein composed of an α chain (36 kDa) and a β subunit (27 kDa) expressed primarily on antigen presenting cells: B cells, dendritic cells, monocytes, macrophages, and thymic epithelial cells. HLA-DR is also expressed on activated T cells. This molecule plays a major role in mediating cellular interactions during antigen presentation to CD4-positive T cells.

552764 Rev. 8
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
552764 Rev.8
Citations & References
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Development References (13)

  1. Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
  2. Dieckmann D, Plottner H, Berchtold S, Berger T, Schuler G. Ex vivo isolation and characterization of CD4(+)CD25(+) T cells with regulatory properties from human blood. J Exp Med. 2001; 193(11):1303-1310. (Biology). View Reference
  3. Herodin F, Thullier P, Garin D, Drouet M. Nonhuman primates are relevant models for research in hematology, immunology and virology. Eur Cytokine Netw. 2005; 16(2):104-116. (Biology). View Reference
  4. Ibisch C, Pradal G, Bach JM, Lieubeau B. Functional canine dendritic cells can be generated in vitro from peripheral blood mononuclear cells and contain a cytoplasmic ultrastructural marker.. J Immunol Methods. 2005; 298(1-2):175-82. (Clone-specific). View Reference
  5. Kitani A, Chua K, Nakamura K, Strober W. Activated self-MHC-reactive T cells have the cytokine phenotype of Th3/T regulatory cell 1 T cells. J Immunol. 2000; 165(2):691-702. (Clone-specific). View Reference
  6. Moran TP, Collier M, McKinnon KP, Davis NL, Johnston RE, Serody JS. A novel viral system for generating antigen-specific T cells. J Immunol. 2008; 175(5):3431-3438. (Clone-specific). View Reference
  7. Pawelec G, Ziegler A, Wernet P. Dissection of human allostimulatory determinants with cloned T cells: stimulation inhibition by monoclonal antibodies TU22, 34, 35, 36, 37, 39, 43, and 58 against distinct human MHC class II molecules. Hum Immunol. 1985; 12(3):165-176. (Biology). View Reference
  8. Pawelec GP, Shaw S, Ziegler A, Muller C, Wernet P. Differential inhibition of HLA-D- or SB-directed secondary lymphoproliferative responses with monoclonal antibodies detecting human Ia-like determinants. J Immunol. 1982; 129(3):1070-1075. (Biology). View Reference
  9. Podolin PL, Bolognese BJ, Carpenter DC, et al. Inhibition of invariant chain processing, antigen-induced proliferative responses, and the development of collagen-induced arthritis and experimental autoimmune encephalomyelitis by a small molecule cysteine protease inhibitor. J Immunol. 2008; 180(12):7989-8003. (Biology). View Reference
  10. Sorg RV, Kogler G, Wernet P. Identification of cord blood dendritic cells as an immature CD11c- population. Blood. 1999; 93(7):2302-2307. (Biology). View Reference
  11. Ziegler A, Heinig J, Muller C, et al. Analysis by sequential immunoprecipitations of the specificities of the monoclonal antibodies TU22,34,35,36,37,39,43,58 and YD1/63.HLK directed against human HLA class II antigens. Immunobiology. 1986; 171(1-2):77-92. (Biology). View Reference
  12. Ziegler A, Uchańska-Ziegler B, Zeuthen J, Wernet P. HLA antigen expression at the single cell level on a K562 X B cell hybrid: an analysis with monoclonal antibodies using bacterial binding assays.. Somatic Cell Genet. 1982; 8(6):775-89. (Biology). View Reference
  13. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
View All (13) View Less
552764 Rev. 8

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.