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Home Q & A Board Biology-Related Homework Help Biochemistry
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yuu yuu
wrote...
13 years ago
Will denaturing sds-page reverse formaldehyde cross-linking?  (5 min at 95C in sample buffer with BME)  Or will the proteins/nucleic acids still be cross-linked on the gel?

Thanks!
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wrote...
#1
Educator
13 years ago
Your wording is somewhat off, yuu. As far as I'm concerned, SDS-page will not reverse (or de-crosslink) the cross-linkings formed by formaldehyde. In principle, heating can reverse the formaldehyde crosslinking. That is why some people just boil their content, but then again boiling (assuming the temperature is 100 C) denatures DNA and proteins. Aldehydes form amid bonds when reacting with primary amino groups, which can be removed by boiling.
wrote...
#2
Staff Member
13 years ago
to add, formaldehyde crosslinks might cause some irreversible damage to proteins, but like ^ said, i've heard that 95 C heating works.
- Master of Science in Biology
- Bachelor of Science
yuu Author
wrote...
#3
13 years ago
Thank you.

So if I heat samples in SDS sample buffer with BME for 5 min at 95... the cross-linking will be reversed?  So it will be individual proteins, not complexes, visible on the gel?
wrote...
#4
Educator
13 years ago
Yes, heat first, then SDS - you will get separation i.e. tetramers become monomers etc.
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