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Material
The DHPHC1 domain of human Vav1 protein has been produced in a bacterial expression system. It contains a mutated amino-acid that mimics tyrosine phosphorylation which is required for interaction with Rac1 (Y174D mutant). It is also 6xHis tagged at its amino terminus for purification purposes. The accession number is AAH13361. The molecular weight of GE05 is approximately 41 kDa. The Vav1 protein is supplied as a white lyophilized powder. Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 4-20% polyacrylamide gradient gel. Vav1 DHPHC1 protein was determined to be approximately 80% pure (see Figure 1).
Reagents
1. Vav1 DHPHC1 protein (Cat. # CS-GE05)
2. Rac1, 2 or 3 protein (Cat. # RC01, CS-RC02 or CS-RC03)
3. Exchange buffer 2 (20 mM Tris-HCl pH 7.5, 50 mM NaCl, 1 mM DTT, 2 mM EDTA, 100 ug/ml BSA, and 0.75 uM Bodipy-FL-GDP), note - make fresh.
4. 50 mM MgCl2in 20 mM Tris-HCl pH 7.5, 50 mM NaCl.
5. 5 mM GTP in 20 mM Tris-HCl pH 7.5, 50 mM NaCl.
Equipment
1. Fluorescence spectrometer (?ex=485nm, ?em=525nm, both with< 15nm bandwidth)
2. Corning 96-well half area plates (Cat # 3686) or other plate with low protein binding surface.
Note: The presented information and documents (Manual, Product Datasheet, Safety Datasheet and Certificate of Analysis) correspond to our latest update and should serve for orientational purpose only. We do not guarantee the topicality. We would kindly ask you to make a request for specific requirements, if necessary.
All products are intended for research use only (RUO). Not for human, veterinary or therapeutic use.
CS-GE05.pdf
CS-GE05-safety-datasheet.pdf
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