CD184 (CXCR4) Monoclonal Antibody (12G5), PE-eFluor™ 610, eBioscience™

Product Specific Information

Description: The 12G5 monoclonal antibody reacts with human CD184 (CXCR4), also termed Fusin, LESTR, or HUMSTR. CD184 is a member of the G-protein-coupled chemokine receptor family with seven membrane-spanning domains, and functions as a coreceptor for X4 HIV-1 entry into CD4+ cells. CD184 is expressed predominantly on naive T cell subsets of peripheral blood and is rapidly upregulated by PHA and IL-2 stimulation. The 12G5 antibody shows partial inhibition of chemotaxis and calcium influx induced by SDF-1 (the natural ligand of CD184), blocks CD4-independent HIV-2 infection, and blocks CD4-dependent infection by some T-tropic HIV-1 isolates.

Applications Reported: This 12G5 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This 12G5 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (1.0 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

PE-eFluor 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red). Please make sure that your instrument is capable of detecting this fluorochrome.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.

Target Information

CXCR4 (Fusin, LESTR, HUMSTR) is a member of the G-protein-coupled chemokine receptor family with seven membrane-spanning domains, and functions as a co-receptor for X4 HIV-1 entry into CD4+ cells. CXCR4 has been proved to be the co-receptor for HIV-2's binding to CD4 through envelope glycoprotein gp 120 and promotes Env-mediated fusion of the virus. In other cases, CXCR4 can even function as the only receptor for HIV-2's binding to the CD4 host cells. CXCR4 is the receptor for the C-X-C chemokine CXCL12/SDF-1 that transduces a signal by increasing intracellular calcium ion levels and enhancing MAPK1/MAPK3 activation. CXCR4 acts as a receptor for extracellular ubiquitin; leading to enhanced intracellular calcium ions and reduced cellular cAMP levels. Also, CXCR4 is involved in hematopoiesis and in cardiac ventricular septum formation, and plays an essential role in vascularization of the gastrointestinal tract, probably by regulating vascular branching and/or remodeling processes in endothelial cells. In the CNS, CXCR4 could mediate hippocampal-neuron survival and is involved in cerebellar development. CXCR4 binds bacterial lipopolysaccharide (LPS) and mediates LPS-induced inflammatory response, including TNF secretion by monocytes. Antibodies to CXCR4 block HIV-1 and HIV-2 fusion and infection of human target cells. The amino-terminal domain and the second extracellular loop of CXCR4 serve as HIV biding sites.