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Visualization: Immuno-Fixation

Dez 17, 2008

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).

Gelreh

The Immunoreaction is done in the CleanPool in 30 ml (100 µl Antibody),
2 hours slightly rocked.

pfeil135
pfeil45
Stainshk

Overnight: Washing out exceed amount of the antibody

autostainklein

Silver staining can be automated with GE's Autostainer

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pH611kurz
Immunfix Silver Stain

Gel: FocusGel 6-11 24S
Visualization: Immuno-Fixation followed by silver staining
(Courtesy of G.Cox, Charlton Memorial Hospital, Swansea)