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