Conference Proceedings

NONREDUCING 2-DIMENSIONAL POLYACRYLAMIDE-GEL ELECTROPHORETIC ANALYSIS OF HUMAN COLONIC PROTEINS

GE REID, H JI, JS EDDES, RL MORITZ, RJ SIMPSON

ELECTROPHORESIS | WILEY | Published : 1995

Abstract

Immunochemical detection of proteins with antigenic determinants that are dependent on the native spatial conformation of the protein can often pose problems with conventional two-dimensional polyacrylamide gel electrophoresis (2-DE). For example, many antigenic determinants are readily destroyed by reducing agents and/or urea, reagents which are critical components of many of the conventional isoelectric focusing and immobilized-pH-gradient (IPG) protocols used in the first electrophoretic dimension. Here we describe the use of commercially available precast 2-DE gels for performing nonreducing/non-urea 2-DE of proteins extracted from the human colon cancer cell line LIM 1215 with 0.3% Trit..

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University of Melbourne Researchers