How to inactivate proteins in organ extracts?

Pow Joshi via methods%40net.bio.net (by pow.joshi At gmail.com)
Fri Dec 8 15:02:22 EST 2006


On 7 Dec 2006 14:03:47 -0800, WS <novalidaddress At nurfuerspam.de> wrote:
> Dear Experts!
>
> How to inactivate proteins in an aqueous organ extract? An attempt to
> digest them with trypsin failed, probably due to trypsin inhibitor from
> blood / serum present in the samples. Heating / boiling comes into my
> mind, but this might kill other unknown substances in the extract.
>
> I would like to tell if a certain effect of this extract is due to a
> protein or something else.
>
> Any ideas?

a tricky problem I must say .........two suggestions/ ideas:

a. you probably could use a combination of proteases .... and pronase
would work well, although that too would'nt cleave all your proteins.
b. you could use some organic solvent, CHCl3/ phenol etc.,  just like
one does when isolating nucleaic acids, and take only the aqueous
phase ...you may have to extract serially .... alternately, you could
use a combination of the two ....
Frankly, when I had to do this sorta experiment, I went to simplest
procedure of heating to about 70-90ºC.

hope that helps to add to the crazy idea database :))
pow



>
> Thanks for your input!
>
> Wolfgang
>
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