Protein sequencing ??!!

Ron Tate rtate at bmb-fs1.biochem.okstate.edu
Tue Dec 10 10:54:23 EST 1996


Adrian Bracken wrote:
> 
> I plan to run total yeast proteins on SDS gels. Certain lanes will contain
> bans not present in other lanes....
>    I am wondering if anybody out there has knowledge about cutting out and
> sequencing such proteins? I would be very grateful if you could reply to
> brackena at tcd.ie. Thanks very much, Adrian Bracken.

Adrian,

I see potential problems with your approach the largest of which is that
you need good resolution of the bands of interest from all the others,
with crude extracts this is difficult to do and may result in
difficulties reading the sequence you get, but its not impossible.  A
rule of thumb I have heard and used is that after you run your gel
transfer proteins to PVDF then stain the PVDF with Coomasie G-250 and
photocopy it, if you can see the photocopied band then you probably have
enough to sequence and you can cut the PVDF band out and put it into the
sequencer.

Good luck and Happy Hunting
-- 
>>>>>>--------------------------->
Ron
Tate                                                                   
Lab of Franklin Leach
Dept. of Biochem. & Molecular Biology
Oklahoma State University 
rtate at bmb-fs1.biochem.okstate.edu
(405) 744-9326
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