Boil (15 mins), Q-Sepharose and Mono-Q
Method based on:
Helps, NR. Street, AJ. Elledge, SJ. And Cohen, PTW. (1994).
“Cloning of the complete coding region for human protein phosphatase inhibitor 2 using the two hybrid system and expression of inhibitor 2 in E. coli”
FEBS letters. 340. 93 - 98.
Enzymatic Assay Format:
Standard phosphatase assay IC50 of 1 nM against PP1 gamma using [ 32P] phosphorylase a as substrate
Calculated molecular mass:
Mono-Isotopic Mass: 23, 001.8 daltons
Average Mass: 23, 016.13 daltons
@Cloning of the complete coding region for human protein phosphatase inhibitor 2 using the two hybrid system and expression of inhibitor 2 in E. coli”
20 mM Ammonium acetate pH 7, 0.1 mM EGTA, 1 mM DTT
Gel Information :
Gel Image 1:
Amino acids M1 – S205 (end) of human Inhibitor 2.
Nde1 and BglII of pCW
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