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PerCP-Cy™5.5 Mouse Anti-Human CD4
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PerCP-Cy™5.5 Mouse Anti-Human CD4
Flow cytometric analysis of CD4 expression on human peripheral blood lymphocytes. Human whole blood was stained with either PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control (Cat. No. 550795; dashed line histogram) or PerCP-Cy™5.5 Mouse Anti-Human CD4 antibody (Cat. No. 566316; solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histogram showing CD4 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Flow cytometric analysis of CD4 expression on human peripheral blood lymphocytes. Human whole blood was stained with either PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control (Cat. No. 550795; dashed line histogram) or PerCP-Cy™5.5 Mouse Anti-Human CD4 antibody (Cat. No. 566316; solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histogram showing CD4 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Pharmingen™
L3T4; Leu3a; T-cell surface antigen T4/Leu-3 ; W3/25 ; CD4 antigen (p55)
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human Peripheral Blood T Cells
Flow cytometry (Routinely Tested)
5 µl
I T38; III T140,496
920
AB_2739678
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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  7. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Cy is a trademark of GE Healthcare.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566923 Rev. 3
Antibody Details
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SK3

The SK3 monoclonal antibody specifically binds to CD4. The CD4 antigen is a 55 kDa type 1 transmembrane glycoprotein that belongs to the immunoglobulin superfamily. It is expressed on T-helper/inducer lymphocytes and monocytes. CD3+CD4+ T cells comprise approximately 28% to 58% of normal peripheral blood lymphocytes. CD4 is also expressed on 80% to 95% of normal thymocytes. The CD4 antigen is present in low density on the cell surface of monocytes and in the cytoplasm of monocytes and macrophages (CD3-CD4+). CD4 serves as a T cell coreceptor for MHC class II antigen recognition and as a receptor for the human immunodeficiency virus (HIV). The SK3 antibody inhibits HIV binding to CD4+ cells.

566923 Rev. 3
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-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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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
566923 Rev.3
Citations & References
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Development References (6)

  1. Bernard A, Boumsell L, Hill C. Joint report of the first international workshop on human leucocyte differentiation antigens by the investigators of the participating laboratories. In: Bernard A, Boumsell L, Dausset J, Milstein C, Schlossman SF, ed. Leucocyte Typing. New York, NY: Springer-Verlag; 1984:9-108.
  2. Engleman EG, Benike CJ, Glickman E, Evans RL. Antibodies to membrane structures that distinguish suppressor/cytotoxic and helper T lymphocyte subpopulations block the mixed leukocyte reaction in man. J Exp Med. 1981; 154(1):193-198. (Clone-specific: Functional assay, Inhibition). View Reference
  3. Evans RL, Wall DW, Platsoucas CD, et al. Thymus-dependent membrane antigens in man: inhibition of cell-mediated lympholysis by monoclonal antibodies to TH2 antigen. Proc Natl Acad Sci U S A. 1981; 78(1):544-548. (Immunogen: Flow cytometry, Inhibition). View Reference
  4. Reichert T, DeBruyere M, Deneys V, et al. Lymphocyte subset reference ranges in adult Caucasians. Clin Immunol Immunopathol. 1991; 60(2):190-208. (Biology). View Reference
  5. Sattentau QJ, Dalgleish AG, Weiss RA, Beverley PC. Epitopes of the CD4 antigen and HIV infection. Science. 1986; 234(4780):1120-1123. (Biology). View Reference
  6. Wood GS, Warner NL, Warnke RA. Anti–Leu-3/T4 antibodies react with cells of monocyte/macrophage and Langerhans lineage. J Immunol. 1983; 131(1):212-216. (Biology). View Reference
View All (6) View Less
566923 Rev. 3

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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.