From: Mario Roederer (Roederer@Beadle.Stanford.EDU)
Date: Thu Dec 19 1996 - 19:51:58 EST
> Apparently, the conjugation with APC can be tricky. For > bright antigens, a good alternative is using Cy5 (kits > from Amersham), or one of the long wavelength BODIPY > dyes from Molecular Probes to make antibody conjugates. Actually, APC conjugations are relatively straightforward. With purified APC on hand, the entire procedure takes about 4-6 hours, requiring 3 quick columns--all on the lab bench. Detailed protocols for APC conjugation, as well as PE, tandems, and small organic fluorescent molecules, can be found at http://cmgm.stanford.edu/~roederer/abcon By the way, Cy5 is a good replacement for APC only if you are using a HeNe laser (or equivalent wavelength). Cy5 is NOT efficiently excited by dye lasers operating near 600 nm (but it does, however, fluoresce some at that excitation wavelength). mr
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