Advantages of preelectrophoretic conjugation of polypeptides with fluorescent dyes

Anal Biochem. 1983 Jul 15;132(2):334-7. doi: 10.1016/0003-2697(83)90016-7.

Abstract

A rapid simple procedure is described for the conjugation of proteins, glycoproteins, and peptides with the fluorescent dye fluorescein isothiocyanate during the time required to polymerize a polyacrylamide gel. Such conjugation does not perturb the electrophoretic mobility of the polypeptides in detergent containing gels. The location of polypeptide . dye conjugate is evident by inspection immediately upon removal of a gel from an electrophoresis cabinet avoiding the time required for postelectrophoretic staining and destaining procedures. The sensitivity of detection of polypeptide . fluorescein conjugates is at least equivalent to that obtained using Coomassie blue.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Electrophoresis, Polyacrylamide Gel / methods
  • Fluorescein-5-isothiocyanate
  • Fluoresceins*
  • Fluorescent Dyes*
  • Glycoproteins / isolation & purification
  • Peptides / isolation & purification*
  • Rosaniline Dyes
  • Thiocyanates*

Substances

  • Fluoresceins
  • Fluorescent Dyes
  • Glycoproteins
  • Peptides
  • Rosaniline Dyes
  • Thiocyanates
  • Fluorescein-5-isothiocyanate
  • coomassie Brilliant Blue