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BV480 Mouse Anti-Human CD4
BV480 Mouse Anti-Human CD4

Flow cytometric analysis of CD4 expression on Rhesus macaque peripheral blood lymphocytes. Rhesus macaque whole blood was stained with either BD Horizon™ BV480 Mouse IgG1, κ Isotype Control (Cat. No. 565652; dashed line histogram) or BD Horizon BV480 Mouse Anti-Human CD4 antibody (Cat. No. 566148/566159; 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 cytometry was performed using a BD LSRFortessa™ Cell Analyzer System.

Flow cytometric analysis of CD4 expression on Rhesus macaque peripheral blood lymphocytes. Rhesus macaque whole blood was stained with either BD Horizon™ BV480 Mouse IgG1, κ Isotype Control (Cat. No. 565652; dashed line histogram) or BD Horizon BV480 Mouse Anti-Human CD4 antibody (Cat. No. 566148/566159; 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 cytometry was performed using a BD LSRFortessa™ Cell Analyzer 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), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse BALB/c IgG1, κ
Human HPB-ALL Cell Line
Flow cytometry (Routinely Tested)
5 µl
AB_2739546
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 BD Horizon BV480 under optimum conditions, and unconjugated antibody and free BD Horizon BV480 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD Optibuild Brilliant reagents) are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The Brilliant Stain Buffer was designed to minimize these interactions.  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).

For Immunofluorescence Applications:

The use of a mounting reagent (eg, ProLong® Gold) is highly recommended to maximize the photostability of BV480.  For confocal microscopy systems, a 440 nm laser is the optimal excitation source and the recommended emission filter is a 485/20 nm bandpass filter.  

For epifluorescence microscopes with broad spectrum excitation sources,  the recommended excitation and emission filters are 445/20 nm and 485/20 nm bandpass filters, respectively.  For specific multicolor imaging applications, the exact filter configurations should be optimized by the end user. For additional instrument/filter configuration information, please visit http://www.bdbiosciences.com/research/cellularimaging.

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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. ProLong® is a registered trademark of Thermo Fisher Scientific, Inc. Waltham, MA.
  9. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566148 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.

The antibody was conjugated to BD Horizon BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set.  BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.

566148 Rev. 2
Format Details
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BV480
The BD Horizon Brilliant Violet™ 480 (BV480) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology fluorochrome has an excitation maximum (Ex Max) of 440-nm and an emission maximum (Em Max) of 479-nm. Driven by BD innovation, BV480 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 480-nm (e.g., a 525/50 bandpass filter). The increased fluorescence intensity of BV480 and narrower emission spectra, make it a good alternative for BV510 or V500. Due to its excitation profile, BV480 will also has less cross-laser excitation with the UV laser, resulting in less spillover into UV channels compared to BV510. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV480
Violet 405 nm
440 nm
479 nm
566148 Rev.2
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. Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
  7. Pitcher CJ, Hagen SI, Walker JM, et al. Development and homeostasis of T cell memory in rhesus macaque. J Immunol. 2002; 168(1):29-43. (Clone-specific: Flow cytometry). View Reference
  8. 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
  9. 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
  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
566148 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.