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Alexa Fluor® 700 Mouse Anti-Human CD4
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Alexa Fluor® 700 Mouse Anti-Human CD4

Flow cytometric analysis of CD4 expression on Rhesus macaque peripheral blood lymphocytes. Rhesus macaque whole blood was stained with Alexa Fluor® 700 mouse anti-Human CD4 antibody (Cat. No. 560836; solid line histogram) or with a Alexa Fluor® 700 Mouse IgG1, κ Isotype Control (Cat. No. 557882; dashed line histogram). The erythrocytes were lysed with BD PharmLyse™ 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.

Flow cytometric analysis of CD4 expression on Rhesus macaque peripheral blood lymphocytes. Rhesus macaque whole blood was stained with Alexa Fluor® 700 mouse anti-Human CD4 antibody (Cat. No. 560836; solid line histogram) or with a Alexa Fluor® 700 Mouse IgG1, κ Isotype Control (Cat. No. 557882; dashed line histogram). The erythrocytes were lysed with BD PharmLyse™ 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 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_10563410
Aqueous buffered solution containing protein stabilizer 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 to Alexa Fluor® 700 under optimum conditions, and unreacted Alexa Fluor® 700 was 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. 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.
  4. Alexa Fluor® 700 has an adsorption maximum of ~700nm and a peak fluorescence emission of ~720nm. Before staining cells with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560836 Rev. 2
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.

560836 Rev. 2
Format Details
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Alexa Fluor™ 700
Alexa Fluor™ 700 dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 697 nm and an emission maximum (Em Max) at 719-nm. Alexa Fluor™ 700 is designed to be excited by the Red (627–640-nm) laser and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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Alexa Fluor™ 700
Red 627-640 nm
697 nm
719 nm
560836 Rev.2
Citations & References
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Development References (12)

  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. 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
  11. 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
  12. 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
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560836 Rev. 2

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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.