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SDS Page heat up and "fry" the gel and protein (Western Blot)


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#1 Podge

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Posted 21 June 2011 - 04:20 PM

Hey,

we recently had this problem 3 or 4 times now in our lab.
We run our SDS Page (4-12% gradient gels, Invitrogen) in an XCell SureLock chamber with MOPS Buffer. We have the box placed in an ice box. Run is 50min @ 200V.

Sometimes it happens that the whole chamber becomes really, really hot and not just handwarm. I noticed that it gets so hot that the gel cassette even deformed! The Western Blots then look really bad and after it happened the first time we throw the gel away immediately.

My theory: somehow the current leaks and therefor the amps have to go up leading to higher Watts(heat).

has anybody seen this before?

Thanks for any suggestions.

Podge

#2 bob1

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Posted 21 June 2011 - 05:05 PM

I have seen it before with the same system. I am not sure what is causing it, but it can usually be prevented by running at a lower voltage for a longer time.

I think it is something to do with the MOPS buffer that is supplied with the kit, maybe a dilution issue or batch preparation (at Invitrogen's end).

#3 Podge

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Posted 23 June 2011 - 08:52 AM

Hey,

the weired thing is though that we use the same MOPS 20x stock since ages. Those bottles last a long time.

But at least somebody else has experienced this aswell.

Thanks.




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