Product Uses Include
Guanosine gamma thio-phosphate (GTPγS), a non-hydrolyzable GTP analog. Upon resuspension with 50 µl de-ionized water you will have a 20 mM stock which is enough for 50 assays of small G-protein activation.
Purity was determined by HPLC. The product is >75% pure.
Biological activity was determined by the ability of the compound to activate RhoA-His protein (Cat. # RH01). Greater than 90% activation is achieved at a concentration of 200 µM BS01.
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Question 1: Can binding of GTPγS to Ras, Arf, and Rho family GTPases be detected with effector proteins conjugated to Sepharose beads?
Answer 1: Yes, GTPγS can be used to create activated Ras, Arf, and Rho family GTPases which can be pulled down with Sepharose beads conjugated to an effector protein specific for the GTPase of interest. The GTPγS should give greater than five-fold increase in activated small G-protein when compared to a GDP bound protein.
Question 2: What percentage of GTPase in an extract will bind to the GTPγS and so constitute an activated small G-protein?
Answer 2: Approximately 5-10% of the total population of Ras, Arf, or Rho family GTPase will bind to the GTPγS, creating an activated GTPase protein.
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