Biotin conjugation

Protocol describing the conjugation of Biotin to an antibody.

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Edited from a protocol kindly provided by  Dr. Guo-Ping Shi

This procedure is a guideline only. Please note that Abcam are unable to guarantee an antibody or peptide if conjugated by the customer.


  1. Make LCB-NHS (long-chain biotin NHS ester) solution with DMSO  (6 mg/ml, 6.5 mM).
  2. Add 10 µl of LCB-NHS into 1 ml of 2 mg/ml Ab and incubate at RT for 50 min.
  3. Terminate the reaction by adding 0.5 ml of 1 M Tris.HCl, pH 8.0.
  4. Dialyse 24-48 hours against 1X PBS.
  5. Add 0.1% (w/v) NaN3 if necessary.

This protocol can also be used for any other peptides and proteins.

Dr. Guo-Ping Shi
Harvard Medical School
Brigham and Women's Hospital
Cardiovascular Medicine
Boston, MA