GTPgammaS: non hydrolysable GTP analog: 100x stock

GTPgammaS: non hydrolysable GTP analog: 100x stock
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Product Uses Include

  • Create constitutively GTP bound GTPases
  • Activate small G-proteins

Material
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
Purity was determined by HPLC. The product is >75% pure.

Biological Activity
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.

For product Datasheets and MSDSs please click on the PDF links below.   For additional information, click on the FAQs tab above or contact our Technical Support department at tservice@cytoskeleton.com

AuthorTitleJournalYearArticle Link
Joseph, Ivor et al.RAB11A and RAB11B control mitotic spindle function in intestinal epithelial progenitor cellsEMBO reports2023ISSN 1469--221X
Morishita, Jun et al.Identification of a small RhoA GTPase inhibitor effective in fission yeast and human cellsOpen Biology2023ISSN 2046-2441
Lin, Benjamin et al.An AMPK phosphoregulated RhoGEF feedback loop tunes cortical flow–driven amoeboid migration in vivoScience Advances2022ISSN 2375-2548
Ozkan, Alper D. et al.Mechanical and chemical activation of GPR68 probed with a genetically encoded fluorescent reporterJournal of Cell Science2021ISSN 1477-9137
Kovalski, Joanna R. et al.The Functional Proximal Proteome of Oncogenic Ras Includes mTORC2Molecular Cell2019ISSN 1097-4164

 

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.

 

 

If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com