Fluorescence behavior of pyrenyl groups bound to the anti-pyrenyl anti
body was studied. The specific binding was found by an energy transfer
from tryptophyl groups of the antibody to the pyrenyl group. The I-1/
I-3 ratio of pyrenyl fluorescence indicated a nonpolar environment of
the bound pyrenyl group, Fluorescence intensity and decay time, howeve
r, were not different from those in solution. Quenching of pyrenyl flu
orescence by external quenchers was suppressed by the antibody when ac
rylamide or more bulky quenchers were used. These results indicate tha
t the pyrenyl groups are not very tightly and not very deeply bound to
the binding site and retain their inherent photophysical properties i
nside the binding site.