APC-Cy™7 Mouse Anti-Human CD195
Clone 2D7/CCR5 (RUO)
- Brand BD Pharmingen™
- Alternative Name CCR5; C-C chemokine receptor type 5; CC-CKR-5; CKR5; CHEMR13
- Vol. Per Test 5 µl
- Isotype Mouse C57BL/6 IgG2a, κ
- Reactivity Human (QC Testing)
Flow cytometry (Routinely Tested)
Immunofluorescence, Functional assay (Tested During Development)
- Immunogen Human CCR5 Transfected Cell Line
- Workshop No. VII 70307
- Entrez Gene ID 1234
- Storage Buffer Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
APC-Cy™7 is a tandem fluorochrome that combines APC and a cyanine dye (Cy7). Special precautions must be taken with APC-Cy7 conjugates, and cells stained with them, to protect the fluorochrome from long-term exposure to light. Some APC-Cy7 conjugates show changes in their emission spectra with prolonged exposure to paraformaldehyde. Fixed cells should be analyzed within 4 hours of fixation in paraformaldehyde or transferred to a paraformaldehyde-free buffer for overnight storage. Due to nearly identical excitation and emission properties but different spillover characteristics, APC-Cy7 and APC-H7 cannot be used simultaneously.
Suggested Companion Products
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.
- 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.
- 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.
- 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).
- 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.
- APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Ficoll-Paque is a trademark of Amersham Biosciences Limited.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/pharmingen/protocols 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-Paque™ separation, freezing, or exposure to cold temperatures prior to staining have been shown to cause a decrease in staining intensity and inconsistent results.