This Immuno-Fixation is followed by the ammoniacal silver staining. Immuno-fixation and silver staining simply done inside the gel. The silver staining starts from step 5, no TCA-fixing and washing necessary. This method combines the sensitivity of the silver-staining with the specificity of the antibody-reaction.
Advantage: Only IgG will be stained, band-sharpness and sensitivity as good as with the general silver-staining. Disadvantage: One overnight step (washing out of the anti-human IgG serum)
The Immunoreaction is done in a special saoking pool in 30 ml (100 ul Antibody), 2 hours slightly rocked
Overnight: Washing out exceed amount of the antibody
Silver staining can be automated with GE's Autostainer
Gel: IEFGel pH 6-11 Visualization: Immuno-Fixation followed by silver staining (Courtesy of G.Cox, Charlton Memorial Hospital, Swansea)