protein sequencing from a western blot

BOMINAAR at chem.rug.nl BOMINAAR at chem.rug.nl
Thu Nov 9 03:57:00 EST 1995


In article <9511081935.AA18583 at umailsrv1.UMD.EDU> Lynne_A_WHITEHEAD at UMAIL.UMD.EDU (lw75) writes:
>From: Lynne_A_WHITEHEAD at UMAIL.UMD.EDU (lw75)
>Subject: protein sequencing from a western blot
>Date: 8 Nov 1995 11:37:12 -0800

>Hi there!
>I am interested in learning more about protein sequencing from western blots.

>1.  is this method reliable?  How much protein is required for successful
>sequencing?

Yes, the method is reliable. In general about 5 pmole for a N-terminus 
sequence of up to 40 aa

>2. can it be done from an SDS-PAGE gel as well as a native PAGE
>blot?

In theory it can, as long as you are sure that what you get on the native 
PAGE is a single species of polypeptide.

>3. does anyone know of any companies with the sequencing apparatus that 
will>process the samples for you?

 I do know one here in the Netherlands, but 
there should be plenty of them in the States.

>Any references would be appreciated!

>Thanks!
>Lynne Whitehead

Good luck Ton



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