How to count labeled protein in SDS-PAGE gel

David J. Meyer, Ph.D. meyerdj at phibred.com
Thu Nov 9 15:05:15 EST 2000


The emission of 35S is not very different from that of C14, so....

David J. Meyer, Ph.D.
Pioneer Hi-Bred International, Inc.

John E. Wiktorowicz, Ph.D. wrote in message ...
>Yes, peroxide at high temp works. Another approach is to incubate the gels
>in ammonium hydroxide at high (~60°C) to hydrolyze the protein and extract
>the radioactivity from the slices. Use the same trick and digest in
>scintillation vials. The ammonium can be evaporated and scintillation fluid
>added after digestion. Forgot to ask, but this assumes you're trying to
>count tritium and C14. Iodine125 of course can be counted directly (gamma
>emmiter), and S35 may be high enough energy to count without extraction.
>Good luck.
>jw
>
>> From: "lxh" <lxh at dartmouth.edu>
>> Organization: Dartmouth College, Hanover, NH, USA
>> Newsgroups: bionet.molbio.proteins,bionet.molbio.methds-reagnts
>> Date: Sat, 21 Oct 2000 18:16:01 -0400
>> Subject: How to count labeled protein in SDS-PAGE gel
>>
>> Does any know a way to count the radioactivity of proteins from a SDS Gel
>> stripe? What is the best way to extract the protein or dissolve the gel?
I
>> assume it is not a good idea to count it directly in a scintilation
counter.
>> But the extraction protocol I 've seen takes too many steps, and I am
afraid
>> I will lost some proteins since I only have several microgram protein per
>> band. Thanks.
>>
>> lxh
>>
>>
>>
>







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