4-((4-Isothiocyanatophenyl)azo)-N,N-dimethylaniline → Erythrosine

Participants

Reaction Steps

Antibody (2 mg) was dissolved in 2 ml of 1% NaHCO3 pH 8.6, and a 20-ul aliquot of a solution of 1 mg/ml 4-dimethylaminoazobenzene-4'-isothiocyanate in dimethylformamide (DMF) added. The mixture was stirred overnight, then desalted on a Sephadex G-25 (coarse) column (1×30 cm). The ultraviolet-and visible spectrum of the conjugate was compared to that of DAB and antibody alone, to determine the degree of substitution. An erythrosin-antibody conjugate was prepared the same way, except the concentration of the erythrosin-isothiocyanate in DMF was 2.5 mg/ml.

Provenance & Evidence