Need a protocol for Native-PAGE - (Jan/07/2010 )
Hi,everyone. Does anyone has a detail protocol for membrane transfer after native-page? Thanks.
-bio01418-
just follow the same protocol that you use for sds-page unless you are separating basic proteins.
also, methanol is not required in the transfer buffer unless you add sds to facilitate transfer.
methanol is still required to activate a pvdf membrane (not for nitrocellulose).
-mdfenko-
mdfenko on Jan 8 2010, 08:30 AM said:
just follow the same protocol that you use for sds-page unless you are separating basic proteins.
also, methanol is not required in the transfer buffer unless you add sds to facilitate transfer.
methanol is still required to activate a pvdf membrane (not for nitrocellulose).
also, methanol is not required in the transfer buffer unless you add sds to facilitate transfer.
methanol is still required to activate a pvdf membrane (not for nitrocellulose).
Thanks，mdfenko. And I do want to separate a basic protein form cell extract, of which the PI is 9.0. Do you have a protocol for this situation?
-bio01418-
you can use the same buffer but put the membrane on the other side
or
you can try a caps buffer (normally used at pH 11, adjusted with naoh, and 10mM working concentration).
-mdfenko-