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BV421 Mouse Anti-Human HLA-ABC
BV421 Mouse Anti-Human HLA-ABC

Two-parameter flow cytometric analysis of HLA-ABC expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon BV421 Mouse anti-Human HLA-ABC antibody (Cat. No. 565332; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric dot plots showing the correlated expression of HLA-ABC (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals were derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Two-parameter flow cytometric analysis of HLA-ABC expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon BV421 Mouse anti-Human HLA-ABC antibody (Cat. No. 565332; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric dot plots showing the correlated expression of HLA-ABC (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals were derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Product Details
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BD Horizon™
Major histocompatibility complex, class I, A,B,C; HLA class I A,B,C
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl
AB_2739189
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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD Optibuild Brilliant reagents) are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  9. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565332 Rev. 2
Antibody Details
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G46-2.6

The Human Leukocyte Antigen (HLA) complex is the human version of the MHC, helping the immune system distinguish the body's own proteins versus those from foreign invaders, such as viruses.  Humans have three main MHC class I genes, known as HLA-A, HLA-B and HLA-C.  Major histocompatibility complex (MHC) class I molecules, which are widely found on the surface of nucleated cells, function by binding peptides and displaying them on the cell surface to cytotoxic T-cells.  Intracellular degradation of cytosolic proteins by the proteasome generates many of the peptides that load MHC class I molecules.  MHC class I may also serve as an inhibitory ligand for natural killer (NK) cell receptors (KIR, Killer Immunoglobulin-like Receptors), which viruses may modulate expression levels for to evade immune detection.  The G46-2.6 monoclonal antibody binds to a monomorphic epitope on the alpha chain of HLA-A, HLA-B and HLA-C.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

565332 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
565332 Rev.2
Citations & References
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Development References (5)

  1. Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
  2. Crisa L, Cirulli V, Ellisman MH, Ishii JK, Elices MJ, Salomon DR. Cell adhesion and migration are regulated at distinct stages of thymic T cell development: the roles of fibronectin, VLA4, and VLA5. J Exp Med. 1996; 184(1):215-228. (Clone-specific: Flow cytometry). View Reference
  3. Kap YS, van Meurs M, van Driel N, et al. A monoclonal antibody selection for immunohistochemical examination of lymphoid tissues from non-human primates. J Histochem Cytochem. 2009; 57(12):1159-1167. (Clone-specific: Immunohistochemistry). View Reference
  4. Koppelman B, Neefjes JJ, de Vries JE, de Waal Malefyt R. Interleukin-10 down-regulates MHC class II alphabeta peptide complexes at the plasma membrane of monocytes by affecting arrival and recycling. Immunity. 1997; (6):861-871. (Clone-specific: Flow cytometry). View Reference
  5. Xia H, Liu H, Zhang G, Zheng Y. Phenotype and function of monocyte-derived dendritic cells from chinese rhesus macaques. Cell Mol Immunol. 2009; 6(3):159-165. (Clone-specific: Flow cytometry). View Reference
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565332 Rev. 2

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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.