By David P. Siderovski
This quantity is an in-depth therapy of G-Protein Signaling, half A, and may hide common equipment of study of RGS protein research, together with Expression and post-translational amendment, Assays of hole task and allosteric regulate, Electrophysiological equipment and RGS-insensitive Ga subunits, Mouse versions of RGS protein motion, equipment of RGS protein inhibition, and G-protein regulators of version organisms.
Table of Contents
-Expression and post-translational modification
-Assays of hole job and allosteric control
-Electrophysiological tools and RGS-insenstitive Ga subunits
-Mouse versions of RGS protein action
-Methods of RGS protein inhibition
-G-protein regulators of version organisms
Read or Download Regulators of G-Protein Signaling, Part A PDF
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Extra info for Regulators of G-Protein Signaling, Part A
COS7 cells were transfected with expression vectors for CXCR5 and RGS1, RGS3, or RGS13. A day after the transfection, cells were treated with 1 g/ml of CXCL13 for 15 min prior to harvest and preparation of cell lysates. An immunoblot of cell lysates with a pERK-specific antibody reveals a reduction in pERK levels in RGS1-, RGS3-, and RGS13-transfected cells as well as in cells treated with pertussis toxin. Levels of expression of RGS1, RGS2, RGS13, and Erk1/2 are shown. (B) CXCL12-induced cell migration.
2003). Human RGS6 gene structure, complex alternative splicing, and role of N terminus and G protein subunit like (GGL) domain in subcellular localization of RGS6 splice variants. J. Biol. Chem. 278, 30261–30271. , Willard, F. , Chasse, S. , Jones, A. , and Siderovski, D. P. (2003). A seven transmembrane RGS protein that modulates plant cell proliferation. Science 301, 1728–1731. Dohlman, H. , and Thorner, J. (1997). RGS proteins and signaling by heterotrimeric G-proteins. J. Biol. Chem. 272, 3871–3874.
After sorting, the cells should be resuspended in RPMI 1640/10% fetal calf serum (FCS) and incubated at 37 for 1 h prior to any signaling or migration assay. 8. Alternatively, cell populations can be obtained by negative selection by magnetic separation. Purify cells from bulk populations by staining cells with specific monoclonal antibodies and negatively selecting those cells, leaving an enriched cell population. 4/1% BSA fraction V) and stain with biotinylated Mab CD4, CD8, GR-1, Mac-1, Ter119, CD11c, and DX5 to remove T cells, macrophages, erythrocytes, natural killer cells, granulocytes, and dendritic cells.