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Multiparameter flow cytometric analysis of CD4 expression on Human peripheral blood leukocyte populations. Human whole blood was stained with either BD Horizon™ RY610 Mouse IgG1, κ Isotype Control (Cat. No. 571151; Left Plot) or BD Horizon™ RY610 Mouse Anti-Human CD4 antibody (Cat. No. 571285/571345; Right Plot). Erythrocytes were lysed with BD FACS Lysing™ Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD4 (or Ig Isotype control staining) versus side light-scatter signals (SSC-A) was derived from gated events with the forward and side light-scatter characteristics of intact leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ Software.
BD Horizon™ RY610 Mouse Anti-Human CD4
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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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Companion Products
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.
Development References (6)
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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
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Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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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.
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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.
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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
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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.
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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.