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      BV421 Mouse Anti-Human CD45RA
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      This product is the replacement for 740083.
      BV421 Mouse Anti-Human CD45RA
      Multiparameter flow cytometric analysis of CD45RA expression on Human peripheral blood leucocyte populations. Human whole blood was stained with BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438) or BD Horizon™ BV421 Mouse Anti-Human CD45RA antibody (Cat. No. 569620; Right Plot) at 0.5 μg/test. Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plots showing the correlated expression of CD45 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      BV421 Mouse Anti-Human CD45RA
      Flow cytometric analysis of CD45RA expression on Rhesus macaque peripheral blood lymphocytes. Rhesus macaque whole blood was stained with BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD45RA antibody (Cat. No. 569620; solid line histogram) at 0.5 μg/test. The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histogram showing CD45RA expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      BV421 Mouse Anti-Human CD45RA BV421 Mouse Anti-Human CD45RA
      Multiparameter flow cytometric analysis of CD45RA expression on Human peripheral blood leucocyte populations. Human whole blood was stained with BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438) or BD Horizon™ BV421 Mouse Anti-Human CD45RA antibody (Cat. No. 569620; Right Plot) at 0.5 μg/test. Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plots showing the correlated expression of CD45 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Flow cytometric analysis of CD45RA expression on Rhesus macaque peripheral blood lymphocytes. Rhesus macaque whole blood was stained with BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD45RA antibody (Cat. No. 569620; solid line histogram) at 0.5 μg/test. The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histogram showing CD45RA expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Horizon™
      CD45R; PTPRC; LCA; Leukocyte common antigen
      Rhesus,Cynomolgus,Baboon (QC Testing), Human (Tested in Development)
      Mouse IgG1, κ
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      5788
      Aqueous buffered solution containing ≤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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      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. 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.
      7. 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.
      8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      9. 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.
      10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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      569620 Rev. 1
      Antibody Details
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      5H9

      The 5H9 monoclonal antibody specifically binds to the human form of the leukocyte common antigen (LCA), CD45RA. This clone also crossreacts with CD45RA expressed by peripheral blood lymphocytes, monocytes and some granulocytes from rhesus macaque, baboon, and cynomolgus monkeys. The observed frequency of CD45RA positive lymphocytes is higher in the non-human primate compared to what is seen with normal human donor lymphocytes.

      569620 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
      569620 Rev.1
      Citations & References
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      Development References (6)

      1. Arbore G, West EE, Rahman J, et al. Complement receptor CD46 co-stimulates optimal human CD8+ T cell effector function via fatty acid metabolism.. Nat Commun. 2018; 9(1):4186. (Clone-specific: Flow cytometry). View Reference
      2. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
      3. Palermo B, Franzese O, Donna CD, et al. Antigen-specificity and DTIC before peptide-vaccination differently shape immune-checkpoint expression pattern, anti-tumor functionality and TCR repertoire in melanoma patients.. Oncoimmunology. 2018; 7(12):e1465163. (Clone-specific: Flow cytometry). View Reference
      4. Reimann KA, Waite BC, Lee-Parritz DE, et al. Use of human leukocyte-specific monoclonal antibodies for clinically immunophenotyping lymphocytes of rhesus monkeys. Cytometry. 1994; 17(1):102-108. (Biology). View Reference
      5. Sopper S, Stahl-Hennig C, Demuth M, Johnston IC, Dorries R, ter Meulen V. Lymphocyte subsets and expression of differentiation markers in blood and lymphoid organs of rhesus monkeys. Cytometry. 1997; 29(4):351-362. (Biology). View Reference
      6. van der Windt DJ, Dons EM, Montoya CL, et al. T-lymphocyte homeostasis and function in infant baboons: implications for transplantation. Transplantation. 2012; 25(2):218-228. (Clone-specific: Flow cytometry). View Reference
      View All (6) View Less
      569620 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.

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