Hello everyone
I want to study GEFx and its binding with GTPase.
Rxn as such:
GEFx = 2uM
GTPase = 1uM
buffer = 10mM tris, 150mM nacl, 5% gly, 2mM bME (buffer same for both above proteins) + 10mM MgCl2
mantGTP =10uM
GTP = 100uM (for exchange )
The experiment is carried in 30*C using Varioskan. with excitation 360nm and emission at 460nm with interval of 2sec for 250 round.
I expect a drop in mant flourophore emission but the kind i am getting is not convincing enough. Kindly suggest me a way to correctly do the experiment or suggest me another method to study interaction.
Yehia A.-G. Mahmoud
Hi Roy
try this link:
https://www.cellbiolabs.com/sites/default/files/69A37112-3048-812A-2EB994285BC8EF13.pdf
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