Cliff, We have done quite a bit of Cy7APC conjugation... you will find detailed protocols in our publication in Cytometry (Roederer et al., Cytometry 24:191, 1996), as well as on the web (www.drmr.com/abcon/). Cy7 can be purchased from Amersham, but only after signing away your life (please complain to them about this; they are being unncessarily restrictive). We use APC (and PE) from ProZyme (www.prozyme.com). You might also contact PharMingen; they have Cy7APC conjugated avidin, and should soon have direct conjugates as well. I have tested their reagent, and it is better than what we make, so they've obviously optimized it quite a bit more than we were willing to do! Note that for detection of Cy7 tandems you will need filters (785/50 and an appropriate dichroic) and you must use a red sensitive (such as a 3896) PMT for sensitivity. If you want more info or some help, you could just drive down peninsula and visit us (only 30 miles away, but our weather is much better!). mr
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