Assays for Ral activation and detection.
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%.
Ral Activation Assay Buffer, 5X:
Two vials, each containing 18ml of 5X Ral Activation Assay Buffer (RAB): 250mM Tris, pH 7.5, 1M NaCl, 5% NP-40, 50mM MgCl2, 2.5mM dithiothreitol.
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 degrees C from the date of shipment.