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protein aggregation (?) in gelatin zymography

gelatin zymography

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#1 BradH



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Posted 04 June 2012 - 10:37 AM


Whenever I run my tissue extracts on the gelatain SDS gel for zymography, I always end up with most of my sample accumulating at the very top of the 8% separating gel (some weak, fuzzy bands typically appears at approximately the expected size range: 60-90 kDa).

I should mention that others in my lab use the same protocol as myself without this problem, and I didn't have this problem when I performed the same experiment at another institution! My standards (recombinant protein and conditioned media from a cell line) run fine. I suspect problems with sample extraction, but again, I used the same sample extraction protocol previously and I didn't have this problem.

Does anyone have any thoughts or suggestions?



#2 mdfenko


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Posted 05 June 2012 - 12:14 PM

how do you prepare your samples? overboiling can cause aggregation.
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