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 Sub-optimal wet transfer buffer? [View Printable]
collmic

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 Send a personal messsage to collmic Reply with a quote from this post Go to the top of the page

I've been using a standard western blot transfer buffer (192mM Glycine, 25mM Tris, 20%MeOH) and within the last few months when I run the apparatus, the power supply maxes out on mA and I can only get about 50V. I used to run my gels at 100V for 1 hour.
Do any of you have any thoughts as to why this is happening? I've tried changing the reagents, but that doesn't seem to do any better....

Thanks.
.........................

Posted Jul 18, 2008, 16:10 PM
R Bishop

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 Send a personal messsage to R Bishop Reply with a quote from this post Go to the top of the page

Collmic,

When this happens to me, I first scrub my apparatus clean with soap and a sponge. Also clean the leads. Salt can build up anywhere on wet transfer apparatus. Ours is a Biorad semi-dry model.

I typically transfer by western blots in Bjerrum Buffer, which is similar but has SDS in it might help. The recipe is in the SSI Protocols section.
Here's the link

http://www.scientistsolutions.com/Science-Protocols_DisplayProtocol_96093856-ead1-45f4-9c59-6f18d9cc9676_85fd2bab-e755-40a0-baa2-b9474e1e2df0.aspx

Rb
.........................

Posted Jul 23, 2008, 18:45 PM
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