Fluorescence from 2AA-labeled glycans and N-NHS-RED-labeled CtUGGTGT24
(A) HPLC elution profiles for the purification of 2AA-labeled glycans obtained from recombinantly expressed HIV gp120. Top panel, black trace: 2AA-labeled glycans purified from cells treated with 10 μM kifunensine (predominantly GlcNAc2Man9, i.e., 2AA-M9 glycan). Bottom panel: 2AA-labeled glycans purified from cells not treated with kifunensine: mostly 2AA-M9 glycan, but containing 2AA-M5, 2AA-M6, 2AA-M7 and 2AA-M8 glycans as well. We call this mixture 2AA-M5-9.
(B) Fluorescence spectra of 5M-8OH-Q, unlabelled CtUGGTGT24 and NT-RED-NHS-labeled CtUGGTGT24. λExcit = 600 nm. Solid and dashed lines refer to samples with or without 5M-8OH-Q, respectively. Gray: 5M-8OH-Q 2.5 mM; green dashed: unlabelled CtUGGTGT24 1.7 μM; green: unlabelled CtUGGTGT24 1.7 μM plus 5M-8OH-Q 2.5 mM; blue dashed: NT-RED-NHS-labeled CtUGGTGT24 1.7 μM; blue: NT-RED-NHS-labeled CtUGGTGT24 1.7 μM plus 5M-8OH-Q 2.5 mM.