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      BV421 Mouse Anti-Human TCR Vβ5.2/5.3
      BV421 Mouse Anti-Human TCR Vβ5.2/5.3
      Two-color flow cytometric analysis of TCR Vβ5.2/5.3 expression on human peripheral blood T lymphocytes. Human peripheral blood cells were stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 561811) and with either BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human TCR Vβ5.2/5.3 antibody (Cat. No. 567971/568023; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A bivariate pseudocolor density plot showing the correlated expression of TCR Vβ5.2/5.3 (or Ig Isotype control staining) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of viable human lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software.
      BV421 Mouse Anti-Human TCR Vβ5.2/5.3
      Two-color flow cytometric analysis of TCR Vβ5.2/5.3 expression on human peripheral blood T lymphocytes. Human peripheral blood cells were stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 561811) and with either BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human TCR Vβ5.2/5.3 antibody (Cat. No. 567971/568023; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A bivariate pseudocolor density plot showing the correlated expression of TCR Vβ5.2/5.3 (or Ig Isotype control staining) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of viable human lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software.
      Product Details
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      BD Horizon™
      TCR Vβ5.2, TRBV5-6, TCRBV5S2; TCR Vβ5.3, TRBV5-5, TCRBV5S3
      Human (QC Testing)
      Mouse IgG2a, κ
      Flow cytometry (Routinely Tested)
      5 µl
      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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.
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      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.

             For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant™ Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant™ Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant™ Stain Buffer for the same length of time as the corresponding multicolor panel. 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).

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      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. 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).
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. An isotype control should be used at the same concentration as the antibody of interest.
      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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Pacific Blue™ is a trademark of Life Technologies Corporation.
      11. 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 https://www.bdbiosciences.com/en-us/support/product-notices.
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      567971 Rev. 1
      Antibody Details
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      MH3-2

      The MH3-2 monoclonal antibody specifically recognizes the human variable beta 5.2 (TCR Vβ5.2) and variable beta 5.3 (TCR Vβ5.3) domains of the beta subunit for the αβ T cell receptor (TCR Vβ5.2/5.3). The TCR Vβ5.2 and TCR Vβ5.3 domains are encoded by the TRBV5-6 (T cell receptor beta variable 5-6; also known as TCRBV5S2) and TRBV5-5 (T cell receptor beta variable 5-5; TCRBV5S3) gene segments of the multimembered Vβ5 subfamily within the TRB (T cell receptor beta locus), respectively. Functional T-cell receptor beta chains (TCR-β) are generated by genomic rearrangement of TCR variable (Vβ), diversity (Dβ), joining (Jβ) and constant (Cβ) region gene segments by precursor T cells. Distinct αβ TCR containing either TCR Vβ5.2 or TCR Vβ5.3 are clonally expressed on subsets of thymocytes or peripheral CD4+ or CD8+ T cells. These TCR β beta chains can heterodimerize with TCR α chains to form αβ TCR that function as signaling T cell receptors for antigen. Human αβ TCR recognize peptides presented by HLA (MHC) molecules and in conjunction with the associated CD3 complex, can transduce intracellular signals that initiate precursor or mature T cell responses. The MH3-2 antibody may be useful for analyzing the frequencies or numbers of TCR Vβ5.2/5.3-positive thymocytes or mature T cells as well as the levels of TCR Vβ5.2/5.3 expressed by these cells. The MH3-2 antibody can be used in research applications such as flow cytometry to help characterize the TCR Vβ repertoires of precursor or mature T cell populations.

      567971 Rev. 1
      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.
      altImg
      BV421
      Violet 405 nm
      407 nm
      423 nm
      567971 Rev.1
      Citations & References
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      View product citations for antibody "567971" on CiteAb

      Development References (7)

      1. Folch G, Jabado-Michaloud J, Lefranc M-P. Reagents monoclonal antibodies: anti-human TRBV, IMGT repertoire, IMGT Web resources. http://www.imgt.org/IMGTrepertoire/Regulation/antibodies/human/TRB/TRBV/Hu_TRBVMab.html. IMGT®. 2020; 4. (Clone-specific: Flow cytometry).
      2. Kabelitz D, Wesch, D. T-Cell Receptor Analysis by Flow Cytometry. In: Sack U, Tarnok A, Rothe G. Sack U, Tarnok A, Rothe G, ed. Basic Principles, Methods and Clinical Applications of Flow Cytometry. New York: Karger, Basel; 2009:200-210. View Reference
      3. Labrecque N, McGrath H, Subramanyam M, Huber BT, Sékaly RP. Human T cells respond to mouse mammary tumor virus-encoded superantigen: V beta restriction and conserved evolutionary features.. J Exp Med. 1993; 177(6):1735-43. (Clone-specific: Flow cytometry). View Reference
      4. Oh S, Terabe M, Pendleton CD, et al. Human CTLs to wild-type and enhanced epitopes of a novel prostate and breast tumor-associated protein, TARP, lyse human breast cancer cells.. Cancer Res. 2004; 64(7):2610-8. (Clone-specific: Flow cytometry). View Reference
      5. Posnett DN, Romagne F, Necker A, Kotzin BL, Sékaly R-P. First human TCR monoclonal antibody workshop. The Immunologist. 1996; 4(1):5-8. (Clone-specific).
      6. Soudeyns H, Champagne P, Holloway CL, et al. Transient T cell receptor beta-chain variable region-specific expansions of CD4+ and CD8+ T cells during the early phase of pediatric human immunodeficiency virus infection: characterization of expanded cell populations by T cell receptor phenotyping.. J Infect Dis. 2000; 181(1):107-20. (Clone-specific: Flow cytometry). View Reference
      7. Vonderheid EC, Boselli CM, Conroy M, et al. Evidence for restricted Vbeta usage in the leukemic phase of cutaneous T cell lymphoma.. J Invest Dermatol. 2005; 124(3):651-61. (Clone-specific: Flow cytometry). View Reference
      View All (7) View Less
      567971 Rev. 1

      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.