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Western Blotting loading control for large proteins


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#1 alchemic

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Posted 30 April 2013 - 04:19 AM

Hi,

For some time I have had a problem with loading contol for my protein, which is ~250kDa.

Running SDS-page gel, I usually run it so long (5% gel) that anything below 170kDa escape my gel and I can't use any of loading control like GAPDH, Actin, tubulin. I work with HeLa/Hek293 T cells.

I know that I can run a separate gel, run it not so long just to make a loading control (detection of protein ~<100kDa) but I would prefer to have a control on the same gel, it is more reliable.

Any suggestions?


Alchemic

#2 mdfenko

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Posted 30 April 2013 - 04:55 AM

run a gradient gel.

Edited by mdfenko, 30 April 2013 - 04:56 AM.

talent does what it can
genius does what it must
i do what i get paid to do

#3 alchemic

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Posted 30 April 2013 - 05:12 AM

Thanks for prompt reply. I will try that.




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