Applications:Non-radioactive Ral Activation Assay Kit can be used to precipitate Ral-A-GTP from cell lysates & detection by a Ral-A specific antibody.
Affinity Precipitation/Western Blot Analysis:Ral-A-GTP was precipitated from A431 cell lysates that had been loaded with GTPγS. The precipitated Ral-A-GTP was detected by Western Blot analysis using anti-Ral A (1:1000).
Quantity:30 assays
Kit Components:Ral BP1 (Ral Assay Reagent), agarose:300ug of protein in 600ul of a 50% agarose slurry in 50mM Tris, pH 7.5, 200mM NaCl, 1% NP40, 1mM PMSF, 1mM aprotinin prior to the addition of glycerol to 50%.
Anti-Ral A (mouse monoclonal IgG2a):One vial containing 75ug of IgG2a in 300ul of 20mM sodium phosphate pH 7.5, 150mM NaCl, 1.5mM sodium azide. 1mg/ml BSA with glycerol added to 50%.
100X GTPS, 10mM:One vial containing 50ul of 10mM GTPS, 100X stock, in sterile water. Non-hydrolyzable analog of GTP. Sufficient to label 5ml of cell lysates.
100X GDP, 100mM:One vial containing 50ul of 100mM GDP, 100X stock, in sterile water. GDP (Guanosine 5'-Diphosphate) for in vitro labeling of G-proteins in the inactive form. Sufficient to label 5ml of cell lysates.
Storage and Stability:Components are stable for 1 year at -20°C from the date of shipment.
