How should I choose an effective protease inhibitor which can can be used for the purification of a range of proteins to be purified by Q-sepharose or Ni-NTA or SP-sepharose?
Hi there,
Generally one uses a cocktail of antiproteases as cell naturally produces diverse proteas activities. The complete tablets from Roche are very popular : http://www.sigmaaldrich.com/catalog/product/roche/04693132001?lang=fr®ion=FR&gclid=CJjCuruUkc4CFegW0wodDZcEaw
Hi there,
Generally one uses a cocktail of antiproteases as cell naturally produces diverse proteas activities. The complete tablets from Roche are very popular : http://www.sigmaaldrich.com/catalog/product/roche/04693132001?lang=fr®ion=FR&gclid=CJjCuruUkc4CFegW0wodDZcEaw
One additional note: use the EDTA-free protease inhibitor cocktail when performing Ni-NTA affinnity chromatography first because the EDTA strips the Ni2+ off the column.
Recommendations by Dominique and Adam are just perfect. Only wanted to emphasize, when using Q- or SP-sepharose (you may use EDTA-containing cocktail to make sure metalloproteinases are also killed.
since your going to histag purification use EDTA free tablets
like
S8830 SIGMASIGMAFAST™ Protease Inhibitor Cocktail Tablets, EDTA-Free
for use in purification of Histidine-tagged proteins
Synonym: Protease Inhibitor Cocktail.
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