| Product Name |
CHO-K1/PAC1/Gα15 Stable Cell Line |
Full Name |
Human Recombinant Vasoactive Intestinal Polypeptide Receptor Stable Cell Line |
|
Species |
Human |
Description |
The Vasoactive intestinal Peptide/Pituitary Adenylate Cyclase Activating Polypeptide (VPAC) receptor is G protein-coupled and has been divided into at least three types: PAC1, VPAC1, and VPAC2. High expression of PAC1 is observed in CNS and the adrenal medulla. Several splice variants of PAC1 result in proteins that differ at the N-terminus and third intracellular loop; these variants differ in their affinities for PACAP and abilities to activate Gq and Gs. This manual describes establishment of a cell line and a protocol of pharmacologically validated human PAC1 GPCR receptor (Genebank Accession Number: NM_003382). |
Freeze Medium |
45% culture medium, 45% FBS, and 10% DMSO |
Culture Medium |
Ham's F12, 10% FBS, 100 μg/ml Hygromycin B, 400 μg/ml G418 |
Storage |
Liquid nitrogen immediately upon delivery |
Application |
Functional assay for PAC1 receptor |
Application Examples |
Figure 1. PACAP-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/PAC1/Gα15 and CHO-K1/Gα15 cells. The cells were loaded with Calcium-4 prior to stimulation with a PAC1 receptor agonist, PACAP. The intracellular calcium change was measured by FlexStation. The relative fluorescent units (RFU) were plotted against the log of the cumulative doses (10-fold dilution) of PACAP (Mean ± SD, n = 2). The EC50 of PACAP on PAC1 co-expressing with Gα15 in CHO-K1 cells was 1.9 nM. The S/B of PACAP on PAC1 co-expressing with Gα15 in CHO-K1 cells was 14. |
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Document-EXAMPLE: 9652_20090423051825.JPG (JPG)
TECHNICAL MANUAL: 9689_20090427235912.PDF (PDF)