APC Mouse Anti-Human CD195
Clone 3A9 (RUO)
- Brand BD Pharmingen™
- Alternative Name CCR-5; Chemokine (C-C motif) receptor 5; CMKBR5; CKR5; CKR-5; CHEMR13
- Vol. Per Test 20 µl
- Isotype Mouse C57BL/6 IgG2a, κ
- Reactivity Human (QC Testing) Rhesus, Cynomolgus (Tested in Development)
Flow cytometry (Routinely Tested)
- Immunogen Human CCR5 Transfected Cell Line
- Workshop No. VII 70309
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
Allophycocyanin (APC), is an accessory photosynthetic pigment found in blue-green algae. Its molecular weight is approximately 105 kDa. APC has six phycocyanobilin chromophores per molecule, which makes it a very bright fluorochrome that is highly suitable for flow cytometry applications. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously
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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 APC under optimum conditions, and unconjugated antibody and free APC were removed.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Ficoll-Paque is a trademark of Amersham Biosciences Limited.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.