Wanted:Peptide quantitiation method

GLAXO SUITE GLAXO.SUITE at Sunderland.Ac.UK
Mon Apr 1 03:18:31 EST 1996


In article <whc-3103961518330001 at macr3-20.welc.cam.ac.uk>, 
whc at mole.bio.cam.ac.uk says...
>
>Does anyone have a reliable method for measuring the concentration of
>short peptides (say 10-30 aa long). The standard Bradford assay to does
>not work on such shorties.
>
>-- 
>Dr.W.H.Colledge
>Physiological Laboratory
>University of Cambridge
>Cambridge CB2 3EG

One fairly reliable way is amino acid analysis. A protocol along these lines 
should suffice.

Use an accuratly determined aliquot of the peptide. Either weighed or 
pipetted from a specified stock solution.

Post hydrolysis dissolution of the sample should be with an accuratly 
measured aliquot of the analysis buffer and standard solution.

The concentrations of the amino acid constituents is reported. So clearly 
from the amount of the hydrolysis stable amino acid- ile, leu, val and the 
like- an accurate peptide content can be found.

I hope this helps.

LEN BELL.
Sunderland university.




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