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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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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web