Hello there. I am getting significant activity from my enzyme extract in recent times. But whenever I try to run a 12 % SDS-PAGE for the extract I end up with absolutely no bands. I tried to run the enzyme with controls like bovine serum albumin and xylanase and I got clear bands. I try to concentrate the enzyme extract (supernantant of course) with acetone. What do you think I should do? Thanks a lot.
If you think your problem is concentration of your enzyme, you can try silver stain or tricine sds page.
Tricine sds-page will not make things better, silver staining will. You can also stain using protocol 3 (Rapid Coomassie blue staining) from chapter 2 (Proteins and proteomics, a laboratory manual. Richard J. Simpson), which is way more sensitive than regular Coomassie staining.
If you do not have access to the manual, send me a message and I will send you a copy of the protocol.
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