Teaching Western Blots with T antigen and p53

 

Theodore Gurney

Tested Studies in Laboratory Teaching, 2005, Volume 26

Abstract

Students perform Western blots with polyacrylamide gel electrophoresis of detergent extracts of SV40-transformed cell lines grown in monolayer culture. The gels are blotted onto nitrocellulose and treated with a monoclonal antibody against SV40 T antigen or against the tumor suppressor p53. Blots are then treated with a secondary antibody (anti-mouse IgG conjugated to horseradish peroxidase), and exposed to revealing chemicals and x-ray film. I give one application: induction of p53 by ultraviolet light.

Keywords:  cell culture techniques, chemiluminescence, Western blot, immunoblot, PAGE, monoclonal antibodies, p53, SV40 T antigen

Bowling Green State University, Ohio (2004)