Inexpensive Biotinylation of Proteins and Antibodies

[Luke]

Inexpensive Biotinylation of Proteins and Antibodies

•   Biotinamidocaproate N-hydroxysuccinimide ester (Sigma, really cheap)
•   DMSO
•   PBS

•   Biotin-labeling buffer pH 7.4
                  0.1M NaCl (5.84g/L)
                  0.1M NaHCO3 (8.4g/L)

1.   Dilute protein to 1mg/mL and dialyze to biotin labeling buffer. (1 to 10,000 final)
2.   Prepare 10mg/mL biotinaxxx solution in DMSO, ie 1mg for 100uL.
3.   Add 10uL of solution for each mg of protein/antibody.
4.   Incubate RT with agitation for 1 hour.
5.   Immediately dialyze sample to PBS or buffer of choice. (1 to 10,000 final)