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Figure 1. Calcium flux in CXCR2-expressing Chem-1 cell line induced by IL-8 and GROα. CXCR2-expressing Chem-1 cells were loaded with Fluo-4 and calc ...
Figure 2. Inhibition of IL-8 and GROα-mediated calcium flux in CXCR2-expressing Chem-1 cells by SB225002. CXCR2 -expressing Chem-1 cells were loaded ...
ChemiSCREEN™ CXCR2 Calcium-Optimized FLIPR Cell Line
Description:
ChemiSCREEN™ CXCR2 Calcium-Optimized FLIPR Cell Line
Trade Name:
ChemiScreen
Background Information:
CXCR2 is a 7-TM G-protein coupled receptor that binds to the chemokines GROalpha, GRObeta, GROgamma, IL-8, ENA-78, NAP-2 and GCP-2 (Olson and Ley, 2002). Neutrophils, mast cells and microvascular endothelial cells express CXCR2, and interactions of CXCR2 with its ligands promotes chemotaxis of these cell types (Heidemann et al., 2003; Nilsson et al., 1999; White et al., 1998). Studies with mice lacking CXCR2 indicate that CXCR2 promotes growth of primary tumors and secondary metastases (Keane et al., 2004), and plays an essential role in hyperoxia-induced lung injury (Sue et al., 2004). In addition, cytomegalovirus encodes a CXCR2-binding chemokine, vCXC-1, that promotes neutrophil migration to infected cells (Penfold et al., 1999). Chemicon's cloned human CXCR2-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant CXCR2 expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between CXCR2 and its ligands.
Application Notes:
Calcium flux assay, ligand binding assays.
Species Reactivity:
Human
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Cell Line Type:
GPCR Cell Lines
Concentration:
2 x 106 cells/mL
Packaging:
2x106 cells/vial
Cell Type:
Chem-1
Gene Symbol:
- IL8RB
- CD182
- CMKAR2
- CXCR-2
- IL8RA
- IL8R2
- CXCR2
- CDw128b
Format:
Cell Line
Presentation:
Cells are frozen at 2 x 106 cells/mL in DMEM/20% fetal bovine serum containing 10% DMSO. Cell line tests negative for mycoplasma.
Quality Assurance:
"SPECIFICATIONS: EC50 for calcium mobilization by IL-8: ~ 7.2 nM; GROα: ~5.6 nM
Signal/noise ratio at GROα Emax: 568
Z' = 0.90 with EC50 concentration of GROα
HOST CELLS: Chem-1, an adherent cell line expressing the promiscuous G-protein, Gα15.
TRANSFECTION: Full-length human CXCR2 cDNA (Accession Number: M73969)
GROWTH MEDIA: DMEM containing 4.5 g/L glucose/10% heat inactivated fetal bovine serum/1x nonessential amino acids/10 mM HEPES/0.25 mg/ml Geneticin (G418) /100 U/ml each penicillin and streptomycin"
Signal/noise ratio at GROα Emax: 568
Z' = 0.90 with EC50 concentration of GROα
HOST CELLS: Chem-1, an adherent cell line expressing the promiscuous G-protein, Gα15.
TRANSFECTION: Full-length human CXCR2 cDNA (Accession Number: M73969)
GROWTH MEDIA: DMEM containing 4.5 g/L glucose/10% heat inactivated fetal bovine serum/1x nonessential amino acids/10 mM HEPES/0.25 mg/ml Geneticin (G418) /100 U/ml each penicillin and streptomycin"
Protein or Enzyme Type:
GPCRs
Species:
Human
Storage Conditions:
Place cells in liquid nitrogen immediately upon receipt. Maintain frozen in liquid nitrogen for up to 5 years. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37ºC water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a T75 flask containing growth media, and replace media after 8-24 h. Cells are typically passaged 1:10 every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.