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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