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BV650 Mouse Anti-Human CD195
BV650 Mouse Anti-Human CD195
Multiparameter flow cytometric analysis of CD195 expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ BV650 Mouse IgG2a, κ Isotype Control (Cat. No. 563417; Left Panel) or BD Horizon BV650 Mouse Anti-Human CD195 antibody (Cat. No. 564999; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202).  Two-parameter flow cytometric contour plots showing the correlated expression of CD195 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Multiparameter flow cytometric analysis of CD195 expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ BV650 Mouse IgG2a, κ Isotype Control (Cat. No. 563417; Left Panel) or BD Horizon BV650 Mouse Anti-Human CD195 antibody (Cat. No. 564999; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202).  Two-parameter flow cytometric contour plots showing the correlated expression of CD195 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Horizon™
CCR-5; Chemokine (C-C motif) receptor 5; CMKBR5; CKR5; CKR-5; CHEMR13
Human (QC Testing), Rhesus, Cynomolgus (Tested in Development)
Mouse C57BL/6 IgG2a, κ
Human CCR5 Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
VII 70309
AB_2739037
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™ BV650 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV650 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. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564999 Rev. 1
Antibody Details
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3A9

The 3A9 monoclonal antibody recognizes CD195, which is also known as the chemokine receptor, CCR5, a seven transmembrane-spanning G protein-associated molecule. The 3A9 antibody also reportedly cross-reacts with human CCR8. Results of epitope mapping and sequence comparison between CCR5 and CCR8 reveals that the first three amino acid residues for these two receptors are identical: MDY (Met-Asp-Tyr). CCR5 belongs to the β-chemokine receptor family. It is expressed on subsets of T lymphocytes, NK cells, monocytes, macrophages, and dendritic cells. CCR5 regulates lymphocyte chemotaxis activation and transendothelial migration during inflammation. It signals a response to at least three chemokines: RANTES and macrophage inflammatory protein-1 (MIP-1) α and β. Additionally, CCR5 has been found to be a co-receptor for macrophage-tropic HIV-1 on CD4+ cells, a characteristic that is important in viral transmission. Reports indicate that individuals who have partial (heterozygous) or complete (homozygous) deletion of the CCR5 allele, demonstrate resistance to HIV infection. CCR5 has been clustered as CD195 in the VIIth HLDA workshop.

The antibody was conjugated to BD Horizon BV650 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm.  BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter).  Due to the excitation and emission characteristics of the acceptor dye, there will be  spillover into the APC and Alexa Fluor® 700 detectors.  However, the spillover can be corrected through compensation as with any other dye combination.

564999 Rev. 1
Format Details
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BV650
The BD Horizon Brilliant Violet™ 650 (BV650) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 406-nm and an acceptor dye with an emission maximum (Em Max) at 649-nm. BV650, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 650-nm (e.g., a 660/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-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.
altImg
BV650
Violet 405 nm
406 nm
649 nm
564999 Rev.1
Citations & References
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Development References (10)

  1. Campbell JJ, Qin S, Unutmaz D, et al. Unique subpopulations of CD56+ NK and NK-T peripheral blood lymphocytes identified by chemokine receptor expression repertoire. J Immunol. 2001; 166(11):6477-6482. (Biology: Flow cytometry). View Reference
  2. Choe H, Farzan M, Sun Y, et al. The beta-chemokine receptors CCR3 and CCR5 facilitate infection by primary HIV-1 isolates. Cell. 1996; 85(7):1135-1148. (Biology). View Reference
  3. Dambra PP, Loria MP, D'Oronzio L, et al. The Cytokine Receptor Panel: Flow cytometry analysis on lymphocytes from neonates, young, aged normal donors, and from patients with HIV infection or AIDS. 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; 2002:269-271.
  4. Deng H, Liu R, Ellmeier W, et al. Identification of a major co-receptor for primary isolates of HIV-1. Nature. 1996; 381(6584):661-666. (Biology). View Reference
  5. Doranz BJ, Rucker J, Yi Y, et al. A dual-tropic primary HIV-1 isolate that uses fusin and the beta-chemokine receptors CKR-5, CKR-3, and CKR-2b as fusion cofactors. Cell. 1996; 85(7):1149-1158. (Biology). View Reference
  6. Hancock WW. Chemokines and the pathogenesis of T cell-dependent immune responses. Am J Pathol. 1996; 148(3):681-684. (Biology). View Reference
  7. Karlsson I, Malleret B, Brochard P, et al. FoxP3+ CD25+ CD8+ T-cell induction during primary simian immunodeficiency virus infection in cynomolgus macaques correlates with low CD4+ T-cell activation and high viral load. J Virol. 2007; 81(24):13444-13455. (Clone-specific: Flow cytometry). View Reference
  8. Raport CJ, Gosling J, Schweickart VL, Gray PW, Charo IF. Molecular cloning and functional characterization of a novel human CC chemokine receptor (CCR5) for RANTES, MIP-1beta, and MIP-1alpha. J Biol Chem. 1996; 271(29):17161-17166. (Biology). View Reference
  9. Rottman JB, Ganley KP, Williams K, Wu L, Mackay CR, Ringler DJ. Cellular localization of the chemokine receptor CCR5. Correlation to cellular targets of HIV-1 infection. Am J Pathol. 1997; 151(5):1341-1351. (Clone-specific: Flow cytometry). View Reference
  10. Wu L, Paxton WA, Kassam N, et al. CCR5 levels and expression pattern correlate with infectability by macrophage-tropic HIV-1, in vitro. J Exp Med. 1997; 185(9):1681-1689. (Immunogen: Flow cytometry, Functional assay, Inhibition, Neutralization). View Reference
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564999 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.