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SDS and transfer buffer......... - (May/28/2006 )

How much SDS is necessary to facilitale better transfer of big proteins e.g > 300kDa? I have been using 0.1% SDS in the transfer buffer, is this enough or can I use abit more?


0.1 % SDS should be enough.
But I would say that even more important than adding SDS to transfer buffer is to use the righ % of gel.

What % are you using? If you are not using a gradient gel, you should make a 4% gel, otherwise I can guarantee you that most of your protein will not be transfered.

For how long and at which voltage you are transfer is also very important.
I would suggest 40V ON, but you need to test it.