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APC-Cy™7 Mouse Anti-Human CD195

BD Pharmingen™ APC-Cy™7 Mouse Anti-Human CD195

Clone 2D7/CCR5

(RUO)
APC-Cy™7 Mouse Anti-Human CD195
Flow cytometric analysis of CD195 expression on human peripheral blood lymphocytes. Whole blood was stained with either APC-Cy™7 Mouse IgG2a, κ Isotype Control (Cat. No. 557751; dashed line histogram) or APC-Cy™7 Mouse Anti-Human CD195 antibody (Cat. No. 557755/560913 ; solid line histogram). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact lymphocytes.
Flow cytometric analysis of CD195 expression on human peripheral blood lymphocytes. Whole blood was stained with either APC-Cy™7 Mouse IgG2a, κ Isotype Control (Cat. No. 557751; dashed line histogram) or APC-Cy™7 Mouse Anti-Human CD195 antibody (Cat. No. 557755/560913 ; solid line histogram). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact lymphocytes.
Product Details
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BD Pharmingen™
CCR5; C-C chemokine receptor type 5; CC-CKR-5; CKR5; CHEMR13
Human (QC Testing)
Mouse C57BL/6 IgG2a, κ
Human CCR5 Transfected Cell Line
Flow cytometry (Routinely Tested), Functional assay, Immunofluorescence (Tested During Development)
5 µl
VII 70307
1234
AB_396861
Aqueous buffered solution containing BSA, 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 with APC-Cy7 under optimum conditions, and unconjugated antibody and free APC-Cy7 were removed.

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-Paque™ 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. 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. 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.
  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. APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate.
  8. APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. Ficoll-Paque is a trademark of Amersham Biosciences Limited.
  11. Cy is a trademark of GE Healthcare.
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
557755 Rev. 11
Antibody Details
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2D7/CCR5

The 2D7/CCR5 monoclonal antibody specifically binds to the human chemokine receptor CCR5, also known as CD195. CCR5 is a seven transmembrane-spanning G protein-associated molecule that belongs to the beta chemokine receptor family and expressed on a subset of T lymphocytes (CD3+CD45RO+CD95+). CCR5 regulates lymphocyte chemotaxis activation and transendothelial migration during inflammation by signaling a response to at least three chemokines: Regulated upon Activation Normal T-cell Expressed and Secreted (RANTES), Macrophage Inflammatory Protein-1 (MIP-1), and Monocyte Chemoattractant Protein 2 (MCP-2).  Additionally, CCR5 has been found to be a coreceptor 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. This antibody has been shown to block ligand and gp120 binding. It is also able to neutralize HIV infection.  

557755 Rev. 11
Citations & References
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Development References (10)

  1. Beavis AJ, Pennline KJ. Allo-7: a new fluorescent tandem dye for use in flow cytometry. Cytometry. 1996; 24(4):390-395. (Biology). 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. 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
  4. 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
  5. 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
  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. Ponath P. Leukosite, Inc. . .
  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. Roederer M, Kantor AB, Parks DR, Herzenberg LA. Cy7PE and Cy7APC: bright new probes for immunofluorescence. Cytometry. 1996; 24(3):191-197. (Biology). 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. (Biology). View Reference
View All (10) View Less
557755 Rev. 11

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