How to enhance PE signal?

From: Jeffrey Rice (rice@aecom.yu.edu)
Date: Thu May 23 2002 - 21:05:47 EST


Hello,
	I'm running into a problem here with my sorting.  I've got a peptide-tetramer
	using biotinylated
peptide with SA-PE.  On our Calibur, I get between half a log and a log difference
between my negatives
and positives, but on our Vantage+DiVa I lose most of that.  I really need to enhance
the PE signal from
my PE positives to improve my sort.
	Now, I know things like Oregon Green labeled anti-FITC exist.  Do any
	FL2-labeled anti-PE
reagents exist?   Or, FL4-labeled anti-APC would be OK too.

	I've tried switching fluorochromes... I get good labeling on the Calibur with
	a direct labeling of the
protein using Alexa Fluor 647 (FL4), but again I lose the separation between my
populations when I try to
sort on the Vantage.  I could use a brighter SA-fluorochrome conjugate for my tetramers,
or a brigher
fluorochome to label the protein.  But I would think that using SA-PE or SA-APC for
tetramers, or Alexa
Fluor 647 for direct labeling, are among the brightest available.

	Any suggestions would be greatly appreciated, as I'm fairly well stuck at
	the moment.

Jeff

----
Jeffrey Rice
rice@aecom.yu.edu

Diamond Lab
Microbiology and Immunology
Albert Einstein College of Medicine
Bronx, NY



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