Conjugation of lectins with fluorochromes: an approach to histochemical double labeling of carbohydrate components.
Methodical investigations on the coupling of lectins (Con A, LcL, WGA, RcA) to tetramethylrhodamine isothiocyan ate (TRITC) are reported. 20-microgram of TRITC per mg of lectin were found to be the optimal amount of TRITC for the conjugation. With this fluorochrome: protein ratio conjugates were produced which resulted in a specific and brilliant fluorescence in tissue staining. The optimally conjugated lectins were separated on DEAE-Sephadex-A 50. Using two different lectins which were conjugated with TRITC or FITC, respectively, a double labeling of different lectin-binding sites in tissue sections was achieved.
D000818:Animals; D001666:Binding Sites, Antibody; D002241:Carbohydrates / Q000032:analysis*; D003106:Colon / Q000032:analysis*; D037102:Lectins; D009206:Myocardium / Q000032:analysis*; D051381:Rats; D012235:Rhodamines; D013194:Staining and Labeling / Q000379:methods*; D014966:Xanthenes
D002241:Carbohydrates; D037102:Lectins; D012235:Rhodamines; D014966:Xanthenes; C009434:tetramethylrhodamine isothiocyanate