How much Protein needed for sequencing?

noone noone at cancer.bham.ac.uk
Thu Jul 16 09:57:13 EST 1998


In article <35aaf5f3.1810987 at news.fu-berlin.de>,
deleteme_cpeter at zedat.fu-berlin.de wrote:

> Hi dear scientific community!
> 
> What is the minimum amount of protein, in micrograms (50kDa) or
> picomoles, on a SDS-PAGE (or western-blotted) which could still be
> reliably sequenced by a state-of-the-art-method? 
> 
> Many thanks in advance............................Christoph
> --
> Christoph Peter
> PhD Student (Biotechnology)
> cpeter at zedat dot fu-berlin dot de
> 
> >Alles wird gut!<(Beate)


Hi Christoph,

if you're talking about Edman degradation, you need about 1 picomol or
more (e.g. on an ABI Procise) in most labs. Thumb rule is that if you can
stain your protein on a PVDF membrane with Coomassie, it's usually enough
for sequencing (if it's not > 200 kD or so). If your protein is already
known you can do Mass-Spec which needs less than that (I think the
EMBL-Labs in Heidelberg offer a commercial service).
The "cracks" in Germany are in Muenchen (Ansorge) and Heidelberg, but they
should have a good sequencing facility at the MDC in Berlin as well.

Hope this helps,

Peter



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