Description
Application
Non-radioactive Rac1 Activation Assay Kit can be used to precipitate Rac1-GTP from cell lysates, & detection by a Rac1 specific monoclonal antibody.
Research Category
Signaling
Biochem/physiol Actions
Predicted to cross-react with all mammalian species
Anti-Rac1, clone 23A8 (mouse monoclonal IgG2b): Human, mouse and rat, other species cross-reactivity unknown. Note: This antibody may be reused 3-5 times for immunoblot analysis. Add 0.05% sodium azide to the blocking buffer containing the diluted antibody. Do not use sodium azides with HRP
Packaging
Kit capacity: 30 assays
Analysis Note
Routinely evaluated by precipitating Rac-GTP from 3T3 cell lysates that had been loaded with GTPγS. The precipitated Rac-GTP was detected by immunoblot analysis using anti-Rac.
Other Notes
Rac1/Cdc42 Assay Reagent (Pak-1 PBD, Magnetic beads): 300 ug of Pak-1 PBD in 150 uL of glutathione magnetic beads, provided as a 50% beads slurry in PBS containing 50% glycerol for a final volume of 300 uL.
Anti-Rac1, clone 23A8: (Part No. 05-389) One vial containing 250 ug of purified IgG2akappa in 250 uL of storage buffer (0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, containing 0.05% sodium azide).
Mg2+ Lysis/Wash Buffer, 5X, (Part No. 20-168).Two vials, each vial contains 18 mL of 125 mM, HEPES, pH 7.5, 750 mM NaCl, 5% Igepal CA-630, 50mM MgCl2, 5 mM EDTA and 10% glycerol
100X GTPγS, 10mM (Cat.# 20-176) One vial containing 50 uL of 10 mM GTPγS, 100X stock, in sterile water. Non-hydrolyzable analog of GTP. Sufficient to label 5 mL of cell lysates
100X GDP, 100 mM, (Part No. 20-177). One vial containing 50 uL of 100 mM GDP, 100X stock, in sterile water. GDP, Guanosine 5’-Diphosphate, type I, for in vitro labeling of G-proteins in the inactive form. Sufficient to label 5ml of cell lysates.
Replaces: SGT425
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
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.
