Detection of "Invisible" Proteins???

ggordon at opal.tufts.edu ggordon at opal.tufts.edu
Mon Oct 24 19:14:25 EST 1994


Greetings, Netters!

	I have an odd situation that we just became aware of.  We have a 
small polypeptide (51 aa) that we have just been able to purify.  We are trying
to separate it by FPLC to determine the form polymeric form (monomer, di-, tri-,
etc.).  Our problem is this...this sequence contains NO AROMATIC AA's...That 
means it can't be detected by UV-light, and therefore can't have its
concentration determined by spectrophotometry.
	This is very frustrating, since we will get large numbers of fractions
and would like to know which ones contain the optimal ammount of protein.

	Does anyone know of a protein concentration method that doesn't 
require aromatic AA's?  We can't really run this on SDS-PAGE and it seems
that the Bradford assay also needs aromatics.  
	If it helps any, the protein is a QIAGEN fusion with a 6-Histidine
tag which binds Nickel.


Thanks in advance!

	Scott Gordon
	Tufts/Sackler School of Biomedical Science 
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