反应详情

An EZ-Link® Sulfo-NHS-LC Biotin stock (Cat: 23224, Thermo Fisher Scientific, Rockford, Ill.) was freshly prepared by adding 300 μL of ice-cold water in a single polypropylene microcentrifuge tube containing 1.0 mg of EZ-LINK® Sulfo-NHS-LC Biotin resulting in a concentration of 5.99 nMole/μL. To a 0.25 mL solution of ANTI-FLAG® M1 antibody (4.041 mg/mL) in a microcentrifuge tube, 11 μL of the freshly prepared EZ-LINK® Sulfo-NHS-LC Biotin water solution was added. This solution was incubated and mixed continuously for 1 hour at room temperature. After the incubation, the antibody was buffer exchanged to a phosphate buffered saline (PBS) containing 37.5 mM KH2PO4, 112.5 mM K2HPO4, 150.6 mM NaCl, 0.10% 2-Methyl-4-isothiazolin-3-one Hydrochloride (MIT), and 7 mM CaCl2. The post modification concentration was 0.975 mg/mL in approximately 1.0 mL.

1. additionmixed continuously for 1 hour at room temperature