Effect of phosphoric acid on a kinase assay
Posted 14 March 2011 - 04:51 AM
I am running some kinase assays. It contains my enzyme, a substrate and some ATP all in a physiological buffer. I am told my the manufacturer that i can halt the reaction by adding phosphoric acid to 0.06%. Can anyone explain exactly what is happening to the ATP in here? Is the acid sequestering the kinase ATP binding site, is there a shift in balance away from ATP to ADP because of the phosphoric acid?
Essentially i need to know the effect so i can determine if it will interferre with a downstream phosphatase reaction.
Biochemistry is not my thing, clearly - i've tried googling and various ewbsites explaining acids etc but have not got a suitable answer.
Any help from anyone would be most appreciated
Posted 14 March 2011 - 08:58 AM
Posted 15 March 2011 - 12:03 PM
Edited by mdfenko, 15 March 2011 - 12:15 PM.
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