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RB780 Mouse Anti-Human HLA-ABC
Product Details
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BD OptiBuild™
HLA class I; HLA class I A,B,C; HLA-A, HLA-B, HLA-C; HLA-A,B,C; MHC class I; MHC-I
Human (Tested in Development)
Mouse BALB/c IgG2a, κ
Human Tonsil Leucocyte Membranes
Flow cytometry (Qualified)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.

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 to the dye under optimum conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  4. Please refer to for technical protocols.
  5. 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.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. 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.
  8. Please refer to to access safety data sheets (SDS).
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at
  10. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  12. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit
755698 Rev. 1
Antibody Details
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The W6/32 monoclonal antibody recognizes a monomorphic epitope expressed on native β2 microglobulin (β2m)-associated Major Histocompatibility Complex (MHC) class I molecules: Human Leucocyte Antigen (HLA)-A, HLA-B, and HLA-C (HLA-ABC). HLA class I molecules are heterodimers comprised of an ~40-45 kDa, highly polymorphic transmembrane α heavy chain, a type I glycoprotein that is noncovalently-associated with an invariant β2-microglobulin (β2m) light chain. The N-terminal extracellular region of the HLA class I heavy chain is comprised of three domains (α1, α2, and α3). The α1 and α2 domains form a closed antigen-binding groove that accommodates 8-10 aa-peptide antigens. β2m non-covalently associates with the α3 heavy chain domain and promotes HLA class I stability. The W6/32 antibody recognizes a conformational epitope on the HLA class I heavy chain. HLA Class I antigens are normally expressed on most nucleated cells. Their expression is upregulated on activated cells or cells responding to various agents including proinflammatory cytokines or mediators. Reduced HLA Class I expression is found on certain virus-infected or tumor cells. HLA class I antigens expressed on thymic epithelial cells are involved in the positive and negative selection of CD8+ T cell precursors which determines their TCR repertoire during T cell maturation. In the periphery, these HLA Class I antigens serve to either present endogenous antigens or cross-present exogenous antigens for the generation of effector and memory CD8+ T cell responses. Target cell HLA Class I antigens can serve as ligands for inhibitory receptors expressed on NK cells and CD8+ T cells and suppress their cytotoxic responses. Human HLA class I molecules play critical roles in cell-mediated immune responses and tumor surveillance as well as tolerance to self-antigens.

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Format Details
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The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
Blue 488 nm
498 nm
781 nm
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Citations & References
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Development References (6)

  1. Barnstable CJ, Bodmer WF, Brown G, et al. Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens-new tools for genetic analysis.. Cell. 1978; 14(1):9-20. (Immunogen: Cytotoxicity, Immunoprecipitation, Radioimmunoassay). View Reference
  2. Margulies DH, Natarajan K, Rossjohn J, McCluskey J. Major Histocompatibility Complex and Its Proteins. In: Paul WE. Paul WE, ed. Fundamental Immunology 7th Edition. Philadelphia: Lippincott Williams & Wilkins; 2013:487-523.
  3. Parham P, Barnstable CJ, Bodmer WF. Use of a monoclonal antibody (W6/32) in structural studies of HLA-A,B,C, antigens.. J Immunol. 1979; 123(1):342-9. (Clone-specific: Immunoaffinity chromatography, Immunoprecipitation, Radioimmunoassay). View Reference
  4. Serrano A, Tanzarella S, Lionello I, et al. Rexpression of HLA class I antigens and restoration of antigen-specific CTL response in melanoma cells following 5-aza-2'-deoxycytidine treatment.. Int J Cancer. 2001; 94(2):243-51. (Clone-specific: Flow cytometry). View Reference
  5. Shields MJ, Ribaudo RK. Mapping of the monoclonal antibody W6/32: sensitivity to the amino terminus of beta2-microglobulin.. Tissue Antigens. 1998; 51(5):567-70. (Clone-specific: Flow cytometry). View Reference
  6. Wooden SL, Kalb SR, Cotter RJ, Soloski MJ. Cutting edge: HLA-E binds a peptide derived from the ATP-binding cassette transporter multidrug resistance-associated protein 7 and inhibits NK cell-mediated lysis.. J Immunol. 2005; 175(3):1383-7. (Clone-specific: Flow cytometry, Functional assay, Inhibition). View Reference
View All (6) View Less
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Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

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