Melittin → Melittin Biotin

Beteiligte Stoffe

Reaktionsschritte

Melittin (1.7 mg) was dissolved in 2 ml of 0.15M sodium phosphate, pH 7.8. To this solution was added 5 mg of N-hydroxysuccinimidobiotin, available commercially from a number of suppliers, and the biotinylation reaction was allowed to proceed at 25° C. for 2 hours. During this time, a large precipitate formed. The precipitate was collected by contrifugation and then dissolved in 1.5 ml of 0.1M sodium acetate buffer, pH 5.6. This sample was then chromatographed on a Sephadex G-25 column (1.5×40 cm) equilibrated in 0.1M sodium acetate, pH 5.6. The melittin-biotin conjugate was identified in the column eluate by monitoring the absorbance at 280 nm. The peak fractions were pooled and stored at 4° C.

Herkunft & Nachweise