BSA test for native PAGE - (Sep/23/2010 )
Hello everybody,
I´m quite unexperienced in the field of native PAGEs. Therefore I want to get my system stabely running before starting with me "real" samples. Has anybody of you experience in performing a native PAGE with BSA as a test-run? How much BSA per sampe would you recommend (in µg)? How does BSA run in he native PAGE? Any other tips for performing native PAGE?
I woud be very grateful to get some tips!!!
the amount of sample depends on the size and thickness of the gel and the width of the wells and the detection method.
i find with reasonably standard size gels that 1-5ug is good with coomassie staining.
since you are running a gel with bsa as the only sample you can run a series of loadings to optimize.
Thank you for your reply!
I will run a gel that is 1.5 mm thick. Have you any experience in blotting native gels? Special transfer buffer or certain conditions which differ from the "normal" wet-blot?