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BV421 Mouse Anti-Human HLA-DR, DP, DQ
BV421 Mouse Anti-Human HLA-DR, DP, DQ
Flow cytometric analysis of HLA-DR, DP, DQ expression on human peripheral blood lymphocytes. Whole blood was stained with either BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; dashed line histogram) or BD Horizon BV421 Mouse Anti-Human HLA-DR, DP, DQ antibody (Cat. No. 564244; solid line histogram). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of HLA-DR, DP, DQ expression on human peripheral blood lymphocytes. Whole blood was stained with either BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; dashed line histogram) or BD Horizon BV421 Mouse Anti-Human HLA-DR, DP, DQ antibody (Cat. No. 564244; solid line histogram). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Horizon™
Tü39; MHC class II HLA-DR, DP, DQ
Human (QC Testing), Rhesus,Cynomolgus,Baboon (Tested in Development)
Mouse IgG2a, κ
Flow cytometry (Routinely Tested)
5 µl
AB_2738697
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

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. 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.
  8. 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.
  9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564244 Rev. 2
Antibody Details
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Tu39

The TU39 monoclonal antibody specifically recognizes human major histocompatibility (MHC) Class II HLA-DR, DP and most DQ antigens. These antigens are encoded by genes within the Human Leukocyte Antigen (HLA) Complex located on chromosome 6. MHC Class II antigens are transmembrane heterodimeric glycoproteins composed of α chain (36 kDa) and β chain (27 kDa) subunits. They are expressed primarily on antigen presenting cells which include dendritic cells, monocytes, macrophages, thymic epithelial cells, and B cells.  They are also expressed on activated T cells. This molecule plays a major role in mediating cellular interactions during antigen presentation to CD4+ T lineage cells. The TU39 antibody is reportedly useful for immunophenotyping as well as functional studies including the inhibition of mixed lymphocyte reactions and antibody-mediated complement fixation on target cells.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.

564244 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
564244 Rev.2
Citations & References
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Development References (4)

  1. Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
  2. 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. (Clone-specific). View Reference
  3. 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. (Clone-specific: Cytotoxicity, Immunoprecipitation, Inhibition). View Reference
  4. 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. (Clone-specific). View Reference
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564244 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.