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Cytochrome C smear in SDS-PAGE

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Cytochrome C smear in SDS-PAGE

Postby NiceOnTheIce » Wed Mar 09, 2011 6:23 pm

I ran an SDS-PAGE for my lab, and in two of the wells I loaded a Cytochrome C/buffer mixture. Both of these lanes resulted in a light smear starting at approx halfway down the gel and ending approx 3/4 of the way down the gel. What would be the reason for the smear? Note: All of my other protein bands ran properly.

Also, I was shown the proper results (using the same technique) and in terms of my results, there should have been one band approx halfway down the gel.

Thanks.
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Postby canalon » Wed Mar 09, 2011 8:49 pm

CytC degradation?
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Postby NiceOnTheIce » Wed Mar 09, 2011 9:12 pm

Thanks for the response, canalon. Cytochrome C degradation makes sense seeing as the smear started where the band should have been, and continued past it. Could you explain to me how degradation might happen?
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Postby canalon » Thu Mar 10, 2011 12:14 am

Nope. Not only because I have no clue about the stability of cytochrome C, but also because I have noclue on how your were treating your samples: origin, extraction method, temperature, when they were extracted and how they were stored. All those and many other things might have impacted your samples. You will have to look at your notebook and at your procedures to figure it out.
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