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

反応式

CN(C1C=CC(N=NC2C=CC(N=C=S)=CC=2)=CC=1)C.[CH:21]1[C:26](N=C=S)=[CH:25][C:24]2[C:30]3([O:50][C:51](=[O:52])[C:23]=2[CH:22]=1)[C:43]1[C:38](=[C:39]([I:46])[C:40]([O-:45])=[C:41]([I:44])[CH:42]=1)[O:37][C:36]1[C:31]3=[CH:32][C:33]([I:49])=[C:34]([O-:48])[C:35]=1[I:47].[Na+].[Na+]>C([O-])(O)=O.[Na+].CN(C)C=O>[CH:21]1[CH:26]=[CH:25][C:24]2[C:30]3([C:31]4[CH:32]=[C:33]([I:49])[C:34]([OH:48])=[C:35]([I:47])[C:36]=4[O:37][C:38]4[C:39]([I:46])=[C:40]([OH:45])[C:41]([I:44])=[CH:42][C:43]3=4)[O:50][C:51](=[O:52])[C:23]=2[CH:22]=1

参加物

実験手順

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.