Which protein seq. lab service??

[Paul N Hengen]

In article <2smuf9$icv at sun4.bham.ac.uk> altabios at bham.ac.uk (John E. Fox) writes:

: I run a core laboratory which among other things, includes protein sequencing.
: Our record is 10 amino acids off 800 femtomole of sample, although this is at
: the extreme edge of the technique when using an ABI 473.  We can work with
: either free proteins or blots on PVDF. Just give the blots a light stain with
: amido black or similar.  We run the sequencing to GLP (Good laboratory
: practice) standards
: 
: Director Dr. John Fox
: tel  UK  021 414 5450
: fax UK  021 414 3376
: Email   altabios at uk.ac.bham

Dear John:

I'm thinking of detecting proteins using a dye such as Nile Red or SYBR Red
from Molecular Probes and then slicing the protein band directly from the gel.
Is it possible to sequence a protein directly from a polyacryamide gel without
blotting onto PVDF? Do you foresee any problems with the red dye blocking the
microsequencing? What is the minimum amount of protein needed for sequencing?

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* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at ncifcrf.gov |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
* Frederick, Maryland 21702-1201 USA              /--------------------------/*
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