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Antibody Labeling
: Biotinylation of Antibody
Biotinylation of Antibody
Author:
Nanci Donacki
Source:
Contributed by Nanci Donacki
Date Added:
Tue May 14 2002
Date Modified:
Thu Apr 29 2004
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.
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