Do Calcium Ions remain bound to proteins after being run through SDS-PAGE?

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BobV
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Do Calcium Ions remain bound to proteins after being run through SDS-PAGE?

I am curious about the effects of calcium binding potentials on several proteins extracted from muscle tissue.  I was wondering if anyone knew if they would remain bound to the proteins through the extraction and 2-D gel seperation.

varsha
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Hi BobV

Hi BobV
I have not worked with muscles or Ca binding proteins myself but this is what I think.
I would think that Ca may remain bound to the protein during extract preparation if you have a non-denaturing condition would preserve the tertiary structure of the protein (with additional calcium provided in the buffer). However as soon as you add a denatuting component such as urea for 2D electrophoresis the protein is going to unfold and lose the bound calcium. You may be able to renature the protein after the 2D run is ove and see if it can bind calcium possibl in a manner similar to in-gel kinase assays.
Hope this is helpful.