Kit Components
1. Anti active Gαs, Mouse Monoclonal Antibody (Catalog No. 26906): One vial – 22 μL (1mg/mL) in PBS, pH 7.4, contained 50% glycerol. This antibody specifically recognizes GTP- Gαs from all vertebrates.
2. Protein A/G Agarose (Catalog No. 30301): One vial – 400 μL of 50% slurry.
3. 5X Assay/Lysis Buffer (Catalog No. 30303): One bottle – 30 mL of 250 mM Tris-HCl, pH 7.4,750 mM NaCl, 5 mM EDTA, 5% Triton X-100.
4. Anti Gαs, Mouse Monoclonal Antibody (Catalog No. 26006): One vial – 22 μL(1 mg/mL) inPBS, pH 7.4,contained 50% glycerol.
5. 100 X GTPγS (Catalog No. 30302): One vial –100 μL at 10 mM, use 5 μL of GTPγS for GTP-labeling of0.5 mL of cell lysate.
6. 100 X GDP (Catalog No. 30304): One vial –100 μL at 100 mM, use 5 μL of GDP for GDP-labeling of 0.5mL of cell lysate.
Storage
Store all kit components at 4oC until their expiration dates. 
Materials Needed but Not Supplied
1. Stimulated and non-stimulated cell lysates
2. Protease inhibitors
3. 4 °C tube rocker or shaker
4. 1 M MgCl2
5. 2X reducing SDS-PAGE sample buffer
6. Electrophoresis and immunoblotting systems
7. Immunoblotting wash buffer such as TBST (10 mM Tris-HCl, pH 7.4, 0.15 M NaCl, 0.05 %Tween-20)
8. Immunoblotting blocking buffer (TBST containing 5 % Non-fat Dry Milk or 3 % BSA)
9. PVDF or nitrocellulose membrane
10. Secondary Antibody
11. ECL Detection Reagents 
Reagent Preparation
? 1X Assay/Lysis Buffer: Mix the 5X Stock briefly and dilute to 1X in deionized water. Just prior to 
usage, add protease inhibitors such as 1 mM PMSF, 10 μg/mL leupeptin, and 10 μg/mL aprotinin.  
Note:
For research use only .