Sigma-Aldrich
cOmplete™ His-Tag Purification Resin
5893682001
Info: Supplier Page
I am interested in studying the molecular mechanism of oncogenic WNT signal transduction to identify potential targets for therapeutic intervention of colon cancer. This experiment aims to purify the His-tagged Cas9 enzyme (that was expressed in E. coli) and utilize it for genome editing of colon cancer cell lines using the ribonucleoprotein (RNP) format. The protein was purified using this kit following the manufacturer's instructions.
Protein purification
E. coli soluble extract
Imidazole concentration was 5 mM during binding and washing steps.
10 gm of clarified extract was incubated with 15 mL of the resin at 4 deg C for 1 h. The resin was washed thrice with 75 mL of wash buffer. Elution was performed with 250 mM Imidazole containing buffer. The final yield was 340 mg of His-tagged Cas9 protein.
N/A
Easy to use.
None.
Good product for His-tagged protein purification and I have regenerated it multiple times without significant loss in binding capacity.