protein elution

u2528082 at ucsvc.ucs.unimelb.edu.au u2528082 at ucsvc.ucs.unimelb.edu.au
Mon Sep 4 18:52:05 EST 1995


We have done quite a lot of gel elution, with variable success.
The easiest option is to lightly stain the band with Coomasie,
then follow the elution of the dye as a marker for protein elution.
Losses are great!! Normally 50%, but it can be more or less.
You can highlight the bavnd with negative staining by KCl, but the 
sensitivity is not great.
If you want to just sequence, then it is more convenient to elute directly
onto sequencing grade PVDF, and localise by Coomasie staining.
I have heard of direct trypsin digestion in the gel, without prior elution.
The resulting peptides are then said to elute more efficiently.
Good elution.
John
U2528082 at hermes.unimelb.edu.au



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