A new method to assign immunodetected spots in the complex two-dimensional electrophoresis pattern

✍ Scribed by Evelyn Zeindl-Eberhart; Peter R. Jungblut; Prof. Hartmut M. Rabes

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 383 KB
• Volume: 18
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.1150180524

✦ Synopsis

Abstract

A new, easy method for the immunodetection of specific antigens in two‐dimensional electrophoresis (2‐DE) is described. Areas of 2‐DE gels containing antigens of interest are electrophoretically transferred to polyvinylidene difluoride membranes, immunostained with specific antibodies using Fast Red or 5‐bromo‐4‐chloro‐3‐indolyl phosphate/nitro blue tetrazolium as detection systems and counterstained with Coomassie Brilliant Blue. In contrast to conventional methods, it is possible to use this procedure to exactly assign immunoreacting proteins on a single blot to their corresponding and surrounding blue‐stained protein spots.