native page trouble!!!

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avantika
avantika's picture
native page trouble!!!

hi!!
i am working with an extracellular  protein which i purified in the following manner::
30% ammoniuom sulphate precipitation and then size exclusion...
when i run my SDS-PAGE..i can see a clear reduction of bands from the crude to 30% cut to the fractions of size exclusion..
however when i run a NATIVE-PAGE..i dont see any bands in the 30% cut...have tried varying my pH of TRIS..i ran a 10% gel initially..i have now reduced it to 8..

what else can i do??where am i going wrong?
 

Rajiv Dua
Rajiv Dua's picture
Try SEC-HPLC....

Try SEC-HPLC....