Prestained standards for Western blotting

Neal Melvin nrmelvin at acs.ucalgary.ca
Wed Jan 6 23:19:55 EST 1999


Can you freeze/thaw tissue samples multiple times without protein degradation??

Annette C. Hollmann wrote:

> In article <770r95$mfq at ds2.acs.ucalgary.ca> Neal Robert Melvin <nrmelvin at ucalgary.ca> writes:
> >Do prestained standards have to be boiled each time you use them?? What
> >about samples?? If I boil a large quantity of sample and freeze it, does
> >it need to be re-boiled for each run?
> >
> Unless you have some unusual solubulity problem with the protein your are
> interested in,
> you do not need to re-boil samples that have already been boiled in SDS
> once. Just thaw, spin down debris, and load your gel(s).
> I have run as many as 20 gels from the same batch of sample, quantitating
> a variety of proteins, and found no detrimental effects.
>
> I don't know what standards you are using, but the Amersham Rainbow
> markers I use actually work better if you don't  boil them at all -
> boiling fades them. If it doesn't say in the product specifications to
> boil the standards, it's probably unnecessary.
>
> Annette




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