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Biotinylation of Antibody

2025-07-15 免疫技术加入收藏

Biotinylation of AntibodyAuthor: Nanci DonackiSource: Contributed by Nanci Donac

Author: Nanci Donacki

Source: Contributed by Nanci Donacki

Materials

0.1M NaHC03 pH9
DMSO
NHS-Biotin (N-Hydroxysuccinimidobiotin, Sigma #H-1759)
PBS

Procedure

Dialyze the sample against carbonate buffer. After dialysis, adjust sample to 1 mg/ml (or less). Prepare 1 mg/ml solution of NHS-Biotin in DMSO. Add NHS-Biotin in DMSO to antibody at desired ratio e.g. for 1:8 ratio add 125 ml of NHS-Biotin to 1mg of Ab Incubate on rotator for 4 hours at room temperature or overnight at 4C. Dialyze against PBS.