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protein degradation - (Nov/17/2010 )

how can we deduce that our protein extract is degradated from western blot images.
i did WB on samples from brain tissue, that was in -20C since 2 or 3 months, and some 1 week.
i saw different profile on my filter after w.b. the signal is lower in the samples kept 3months in -20C. morever, there are some bands at the top of the filter. but may be this is due to aspecifics because of secondary antibody..
if u have ideas about this topic, thanks for your interaction

-luciana-

the bands at the top of the membrane may be due to aggregation of the antigen. this may account for the reduced signal at the normal migration distance.

-mdfenko-

If the protein is degraded, you will see bands (or smears) at smaller molecular weights

-kfunk106-

its what i see in my western!
smears in below!! so, i think its my samples which are degradated.

-luciana-