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562658__9_BV605_CD4_TDS.png

Flow cytometric analysis of CD4 expression on Human peripheral blood lymphocytes. Human whole blood was stained with the BD Horizon™ BV605 Mouse Anti-Human CD4 antibody (Cat. No. 562658/562659; solid line histogram) or with BD Horizon™ BV605 Mouse IgG1, κ Isotype Control (Cat. No. 562652; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

562658__9_BV605_CD4_TDS.png Orange-Cap_Violet.png

Flow cytometric analysis of CD4 expression on Human peripheral blood lymphocytes. Human whole blood was stained with the BD Horizon™ BV605 Mouse Anti-Human CD4 antibody (Cat. No. 562658/562659; solid line histogram) or with BD Horizon™ BV605 Mouse IgG1, κ Isotype Control (Cat. No. 562652; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Horizon™
L3T4; T-cell surface antigen T4/Leu-3; W3/25; CD4 antigen (p55)
Human (QC Testing)
Mouse IgG1, κ
PHA-stimulated Human Peripheral Blood Mononuclear Cells
Flow cytometry (Routinely Tested)
5 µl
IV T114; VI 6T-079
920
AB_2744420
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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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 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. 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).
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
  8. 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.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. For U.S. patents that may apply, see bd.com/patents.
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562659 Rev. 6
Antibody Details
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RPA-T4

The RPA-T4 monoclonal antibody specifically binds to CD4, a 59 kDa single-chain transmembrane glycoprotein that is expressed on T-helper/inducer cell populations. CD4 is also expressed on thymocyte subsets and at lower levels on monocytes and macrophages. CD4 functions as a co-receptor in MHC class II-restricted antigen-induced T cell activation and as a receptor for human immunodeficiency viruses (HIV). This antibody binds to the D1 domain (CDR1 and CDR3 epitopes) of the CD4 antigen and reacts with approximately 80% of thymocytes and 45% of peripheral blood lymphocytes. RPA-T4 is capable of blocking HIV-1, gp120, and inhibits syncytium formation.

562659 Rev. 6
Format Details
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BV605
The BD Horizon Brilliant Violet™ 605 (BV605) dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 605-nm. BV605, driven by BD innovation, is designed to be excited by the Violet laser (405-nm) and detected using an optical filter centered near 610-nm (e.g., a 610/20-nm bandpass filter). The acceptor dye can be excited by the Yellow-Green (561-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV605
407 nm
605 nm
562659 Rev.6
Citations & References
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View product citations for antibody "562659" on CiteAb

Development References (6)

  1. Ho TH, Pfeffer K, Weiss GJ, Ruiz Y, Lake DF. Identification of a CD4+ T cell line with Treg-like activity.. Hum Immunol. 2022; 83(4):281-294. (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. Moebius U. Cluster report: CD4. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:314-330.
  4. Piatier-Tonneau D. CD4 workshop panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:49-54.
  5. Pucino V, Certo M, Bulusu V, et al. Lactate Buildup at the Site of Chronic Inflammation Promotes Disease by Inducing CD4+ T Cell Metabolic Rewiring.. Cell Metab. 2019; 30(6):1055-1074.e8. (Clone-specific: Flow cytometry). View Reference
  6. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
View All (6) View Less
562659 Rev. 6

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.