Calman, Our first pass at Zenon reagents looks interesting. We stained cells for the intracellular phospho-epitope marked by the mpm2 mAb - which marks mitotic cells. We didn't clean the reaction up by removing free Zenon reagent, however, we did use a subsaturating level (about 1/2 max). The distance between mitotic and G2 cells (median to median) was better for the A647-Zenon reagent than for primary labeled FITC-mpm2 by a small amount. However, the CVs for the Zenon labeled cells were greater - more noticable in the dim G2 cells than the bright M cells. While I don't understand the CV business yet, I am excited about the potential of these reagents. jake -- James W. Jacobberger, PhD Professor, Div. General Medical Sciences Case Western Reserve University, Cancer Center 10900 Euclid Ave.(BUT FOR COURIER SERVICES USE: 2109 Adelbert Rd. Cleveland, OH 44106-4944 ph: 216-368-4645 web site: http://josephine.cwru.edu Wednesday, May 15, 2002, 6:15:15 PM, you wrote: CP> I was a bit doubtful about the hype (may make directly labeled mAbs CP> obsolete), but a week after seeing Richard Haugland's posting about Zenon CP> Fab labeling kits I suddenly faced a situation perfectly suited for their CP> use. We were faced with 4 steps staining protocol, but the Zenon reagent CP> reduced this to one step. Basically these are goat anti-mouse Fab fragments CP> that allow quick 10 minute labeling of primary mAbs; a picture is worth a CP> thousand words (http://www.probes.com/products/zenon/zenon.html). CP> We have been using the Zenon APC reagent for intracellular staining of CP> eosinophils for the past 3 weeks and have been getting very good results CP> with low non-specific binding and bright signal. The Alexa 647 seems to bind CP> eosinophils non-specifically, but this is likely due to the eosinophil, as CP> FITC also binds eos. CP> Many cutting edge mAbs are simply not commercially available as flurochrome CP> conjugates and certainly not as exotic fluorochrome conjugates. This systems CP> allows one a lot more flexibility in experimental design. CP> Not meaning to be overly promotional, but I am interested in other's CP> experiences with or thoughts about these reagents and their strengths and CP> limitations. I'm sure many folks out there could benefit from such a CP> discussion. CP> Calman >> _______________________ >> Calman Prussin >> Laboratory of Allergic Diseases >> NIAID/ National Institutes of Health >>
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