Rac1 Activation Magnetic Beads Pulldown Assay

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

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

Kit capacity: 30 assays

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.

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

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