Nonreducing two‐dimensional polyacrylamide gel electrophoretic analysis of human colonic proteins

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..