Catalog # CK6500


CXCR4 Phospho-Regulation

Antibody Sampler Kit

$395

The chemokine receptor CXCR4 is a widely expressed G protein-coupled receptor required for development, hematopoiesis, organogenesis, and vascularization. In disease, CXCR4 has been implicated in WHIM syndrome, HIV, and cancers. Regulation of CXCR4 function occurs through phosphorylation at multiple sites in the C-terminal region. These sites have been shown to be phosphorylated after CXCL12 stimulation, and involve several kinases, such as PKC and GRK kinases. After CXCL12 stimulation of HEK293 cells, Ser-324 and Ser-325 become phosphorylated by PKC and GRK6, while Ser-330 and Ser-339 are phosphorylated by only GRK6. In human astroglia cells, Ser-324 and Ser-325 are rapidly phosphorylated in endogenous CXCR4, while Ser-330 was phosphorylated with slower kinetics. In addition, arrestin binding to CXCR4 is driven by this phosphorylation of far C-terminal residues. Thus, site-specific phosphorylation of CXCR4 may be regulated by multiple kinases lead to complex regulation of CXCR4 signaling.

References
Orsini, M.J. et al. (1999) J Biol Chem. 274(43):31076.
Orsini, M.J. et al. (2000) J Biol Chem. 275(33):25876.
Bhandari, D. et al. (2009) Mol Biol Cell. 20:1324.
Busillo, J. M. et al. (2010) J Biol Chem. 285(10):7805.
Rabbit polyclonal antibodies are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.



*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

CP4251 Saint-Georges, S. et al. (2016) Oncotarget. 20167(27):41031. WB: human leukemia, IgM activated
CP4211 Lu, Y. et al.(2014) Stem Cells Dev. 23(15):1755. ICC: chicken embryonic fibroblasts
CP4251 Yang, S. et al. (2013) Development. 140(22):4554. IHC: mouse midbrain
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
CP4251 Meechan, D. et al. (2012) PNAS. 109(45):18601. WB: mouse interneurons
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
CP4251 Panneerselvam, J. et al. (2105)PLoS One. 10(3):e0122439 WB: human lung cancer cells

Western blot analysis of human Jurkat cells treated with 100 nM calyculin A for 30 min. then the blots were untreated (-) or treated (+) with lambda phosphatase. The blots were probed with rabbit polyclonals anti-CXCR4 (Ser-324/Ser-325) (left panel) or anti-CXCR4 (a.a. 322-329) (right panel).

Immunocytochemical labeling of CXCR4 in chick pluripotent cells. The cells were labeled with rabbit polyclonal CXCR4 (S324/S325) antibody (CP4251), then detected using appropriate secondary antibody (Red). (Image provided by Dr. Yangqing Lu at the Regenerative Bioscience Center, University of Georgia).



This kit contains:

CATALOG #DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
CXCR4 (a.a. 332-329) Rabbit pAb50 μlWB, E, ICCHu, Rt, Ms, Ck45
CXCR4 (a.a. 328-338) Rabbit pAb50 μlWB, E, ICCHu, Rt, Ms45
CXCR4 (Ser-324/Ser-325), phospho-specific Rabbit pAb 50 μlWB, E, ICC, IHCHu, Rt, Ms, Ck45
Anti-Rabbit Ig Light Chain Specific:HRP Mouse mAb100 μlWB, E, ICC, IHCRb
KIT SUMMARY

The CXCR4 phospho-regulation antibody sampler kit can be used to examine the level of dual phosphorylation of Ser-324/Ser-325 relative to the total CXCR4 expression level in western blot, ELISA, and immunocytochemistry.