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Biotinylation of Antibody

Author: Nanci Donacki
Source: Contributed by Nanci Donacki
Date Added: Tue May 14 2002
Date Modified: Thu Apr 29 2004

Materials

Procedure

  1. Dialyze the sample against carbonate buffer.
  2. After dialysis, adjust sample to 1 mg/ml (or less).
  3. Prepare 1 mg/ml solution of NHS-Biotin in DMSO.
  4. 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
  5. Incubate on rotator for 4 hours at room temperature or overnight at 4C.
  6. Dialyze against PBS.
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