Not available outside of the UK & Ireland.

Components

Rac/cdc42 Assay Reagent (PAK-1 PBD, agarose), Catalog # 14-325 :One vial containing 300 µg of PAK-1 PBD bound to 150 µL of glutathione agarose beads, provided as a 50% slurry in 20mM PBS, pH 7.4, containing 50% glycerol for a final volume of 300 µL.Anti-Rac1, clone 23A8, Catalog #05-389: One vial containing 250 µg of protein G purified mouse IgG_2b in 250 µL of storage buffer (0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, containing 0.05% sodium azide).Anti-cdc42, (mouse monoclonal IgG₁) , Catalog # 05-542: One vial containing 50 µg of purified mouse IgG₁ in 200 µL of 50% storage buffer (20 mM sodium phosphate, pH 7.5, 150 mM NaCl, 1.5 mM sodium azide containing, 1 mg/mL BSA) and 50% glycerol.Mg2+ Lysis/Wash Buffer, 5X, Catalog # 20-168 : Two vials, each vial containing 18 mL of 5X MLB: 125 mM HEPES, pH 7.5, 750 mM NaCl, 5% Igepal CA-630, 50 mM MgCl₂, 5 mM EDTA and 10% glycerol.100X GTPγS, 10mM, Catalog # 20-176: One vial containing 50 µL of 10 mM GTPγS, 100X stock, in 50 mM Tris-HCl, pH 7.8, non-hydrolyzable analog of GTP. Sufficient to label 5 mL of cell lysates.100X GDP, 100mM, Catalog # 20-177: One vial containing 50 µL of 100 mM GDP, 100X stock, in 50 mM Tris-HCl, pH 7.8. GDP (Guanosine 5′-Diphosphate) for in vitro labeling of G-proteins in the inactive form. Sufficient to label 5ml of cell lysates.

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.

General description

The Rac1/Cdc42 Activation Assay (Cat. No. 17-441) provides an effective method for detecting Rac and Cdc42 activity in cell lysates. This assay uses the downstream effector of Rac/Cdc42, p21-activated protein kinase (PAK1), to isolated the active GTP-bound form of Rac/Cdc42 from the sample. The p21 binding domain (PBD) of PAK1 is expressed as a GST-fusion protein and coupled to agarose beads. After precipitation, an immunoblot is performed and the activated Rac/Cdc42 is detected with specific monoclonal antibodies, followed by HRP-conjugated secondary antibody and ECL reagent.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Quality

Routinely evaluated by precipitating Rac1-GTP and cdc42-GTP from 3T3/A31 and HeLa cell lysates that had been loaded with GTPγS(lysates were loaded in vitro with GTPγS for cdc42-GTP). The precipitated Rac1-GTP and cdc42-GTP was detected by immunoblot analysis using anti-Rac1 (1 µg/ml).

Specificity

Species Cross-reactivity: Human and mouse. Predicted to cross-react with all mammalian species.
Anti-Rac1, clone 23A8: Species Cross-reactivity: Human, mouse and rat. Other species cross-reactivity unknown.
Anti-cdc42 Species Cross-reactivity: Human, mouse, rat and dog. Other species cross-reactivity unknown.

Storage and Stability

1 year at -20°C from date of shipment