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BSA test for native PAGE - (Sep/23/2010 )

Hello everybody,

Im 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 g)? How does BSA run in he native PAGE? Any other tips for performing native PAGE?
I woud be very grateful to get some tips!!!

-Janine-

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.

-mdfenko-

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?

-Janine-