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PE-Cy™7 Mouse Anti-Human CD4
PE-Cy™7 Mouse Anti-Human CD4

Flow cytometric analysis of CD4 on lysed whole blood.  Human (left panel) or rhesus macaque (right panel) lysed whole blood was stained with the

PE-Cy™7 Mouse Anti-Human CD4 antibody (Cat. No. 560644; unshaded histogram) or with a PE-Cy™7 Mouse IgG1, κ isotype control (Cat. No. 557872; shaded histogram).  Histograms were derived from gated events based on light scattering characteristics for lymphocytes. Flow cytometry was performed on a

BD™ LSR II flow cytometry system.

Flow cytometric analysis of CD4 on lysed whole blood.  Human (left panel) or rhesus macaque (right panel) lysed whole blood was stained with the

PE-Cy™7 Mouse Anti-Human CD4 antibody (Cat. No. 560644; unshaded histogram) or with a PE-Cy™7 Mouse IgG1, κ isotype control (Cat. No. 557872; shaded histogram).  Histograms were derived from gated events based on light scattering characteristics for lymphocytes. Flow cytometry was performed on a

BD™ LSR II flow cytometry system.

Product Details
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BD Pharmingen™
L3T4 ; T-cell surface antigen T4/Leu-3; W3/25 ; CD4 antigen (p55)
Rhesus, Cynomolgus, Baboon (QC Testing), Human (Tested in Development)
Mouse BALB/c IgG1, κ
Human HPB-ALL Cell Line
Flow cytometry (Routinely Tested)
5 µl
AB_1727474
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 PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed.

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. PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. 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 PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  6. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
  7. 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.
  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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560644 Rev. 3
Antibody Details
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L200

The L200 monoclonal antibody specifically binds to the human form of the 56 kDa transmembrane glycoprotein, CD4, which is present on the T-helper/inducer subset of normal human donor peripheral blood lymphocytes. The L200 antibody also cross-reacts with a subset of CD3-positive peripheral blood lymphocytes, but not monocytes, of both Rhesus and Cynomolgus Macaque monkeys. Cross-reactivity on both lymphocytes and monocytes (weak) from baboons is also observed. CD4 distribution on lymphocytes is similar for both human and monkey cells, with the majority of CD4-positive lymphocytes being CD8-negative and lacking reactivity with antibodies to B- or NK-cell markers.

560644 Rev. 3
Format Details
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PE-Cy7
PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-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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PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
560644 Rev.3
Citations & References
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Development References (13)

  1. Attanasio R, Dilley D, Buck D, et al. Structural characterization of a cross-reactive idiotype shared by monoclonal antibodies specific for the human CD4 molecule. J Biol Chem. 1991; 266(22):14611-14619. (Biology). View Reference
  2. Bleavins MR, Brott DA, Alvey JD, de la Iglesia FA. Flow cytometric characterization of lymphocyte subpopulations in the cynomolgus monkey (Macaca fascicularis). Vet Immunol Immunopathol. 1993; 37(1):1-13. (Biology). View Reference
  3. Giorgi JV, Hultin LE, Desrosiers RC. The immunopathogenesis of retroviral diseases: no immunophenotypic alterations in T, B, and NK cell subsets in SIVmac239-challenged rhesus macaques protected by SIV delta nef vaccination. J Med Primatol. 1996; 25(3):186-191. (Biology). View Reference
  4. Indzhiia LV, Yakovleva LA, Overbaugh J, et al. Baboon T cell lymphomas expressing the B cell-associated surface proteins CD40 and Bgp95. J Clin Invest. 1992; 12(3):225-236. (Biology). View Reference
  5. Jacobsen CN, Aasted B, Broe MK, Petersen JL. Reactivities of 20 anti-human monoclonal antibodies with leucocytes from ten different animal species. Vet Immunol Immunopathol. 1993; 39(4):461-466. (Biology). View Reference
  6. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  7. Powell JD, McClure HM, Anderson D, Fultz PN, Sell KW, Ahmed-Ansari A. Phenotypic and functional differences in NK and LAK cells in the peripheral blood of sooty mangabeys and rhesus macaques. Cell Immunol. 1989; 124(1):107-118. (Biology). View Reference
  8. Savary CA, Lotzova E, Jackson HJ, Jardine JH, Ang KK. Analysis of interleukin-2-activated killer cells of rhesus monkeys: striking resemblance to the human system. J Leukoc Biol. 1993; 54(4):307-313. (Biology). View Reference
  9. 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.
  10. 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.
  11. Tryphonas H, Lacroix F, Hayward S, Izaguirre C, Parenteau M, Fournier J. Cell surface marker evaluation of infant Macaca monkey leukocytes in peripheral whole blood using simultaneous dual-color immunophenotypic analysis. J Med Primatol. 1996; 25(2):89-105. (Biology). View Reference
  12. Verdier F, Aujoulat M, Condevaux F, Descotes J. Determination of lymphocyte subsets and cytokine levels in cynomolgus monkeys. Toxicology. 1995; 105(1):81-90. (Biology). View Reference
  13. Wilson AD, Shooshtari M, Finerty S, Watkins P, Morgan AJ. Selection of monoclonal antibodies for the identification of lymphocyte surface antigens in the New World primate Saguinus oedipus oedipus (cotton top tamarin). J Immunol Methods. 1995; 178(2):195-200. (Biology). View Reference
View All (13) View Less
560644 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.