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Purified Mouse Anti-Human CD195
Purified Mouse Anti-Human CD195
Flow cytometric analysis of CD195 expression on human peripheral blood lymphocytes. Whole blood was stained with either Purified Mouse IgG2a, κ Isotype Control (Cat. No. 555571; dashed line histogram) or Purified Mouse Anti-Human CD195 (Cat. No. 556041; solid line histogram). Secondary staining was carried out with FITC Goat Anti-Mouse IgG/IgM (Cat. No. 555988) and erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Fluorescent histograms were derived from gated events with the side and forward light-scatter characteristics of viable lymphocytes.
Flow cytometric analysis of CD195 expression on human peripheral blood lymphocytes. Whole blood was stained with either Purified Mouse IgG2a, κ Isotype Control (Cat. No. 555571; dashed line histogram) or Purified Mouse Anti-Human CD195 (Cat. No. 556041; solid line histogram). Secondary staining was carried out with FITC Goat Anti-Mouse IgG/IgM (Cat. No. 555988) and erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Fluorescent histograms were derived from gated events with the side and forward light-scatter characteristics of viable lymphocytes.
Product Details
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BD Pharmingen™
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), Fluorescence microscopy (Tested During Development)
0.5 mg/ml
VII 70309
AB_396312
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Immunophenotyping studies of chemokine receptors need to be performed on freshly collected whole blood (<24 Hrs). Incubation with the antibody should be done at room temperature in the dark. Cellular manipulation, such as Ficoll™ separation, freezing, or exposure to cold temperatures prior to staining have been shown to cause a decrease in staining intensity and inconsistent results.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. Ficoll-Paque is a trademark of Amersham Biosciences Limited.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
556041 Rev. 8
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.

556041 Rev. 8
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
556041 Rev.8
Citations & References
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Development References (8)

  1. 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
  2. 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
  3. 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
  4. Dragic T, Litwin V, Allaway GP, et al. HIV-1 entry into CD4+ cells is mediated by the chemokine receptor CC-CKR-5. Nature. 1996; 381(6584):667-673. (Biology). View Reference
  5. Hancock WW. Chemokines and the pathogenesis of T cell-dependent immune responses. Am J Pathol. 1996; 148(3):681-684. (Biology). View Reference
  6. 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
  7. 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. (Biology). View Reference
  8. 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. (Biology). View Reference
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556041 Rev. 8

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