EDC Impressum & DSE

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Visualization: General Silver Staining

A special ammoniacal silver-staining is done. This staining is the most IgG-sensitive silver staining recipe.

Advantage: Oligoclonal bands show up without any loss of band-sharpness.
Disadvantage: Not only IgG, all other proteins get stained.

This staining is not IgG-selective, all proteins are stained.
The reason why this visualization procedure gives enough information about the different the Oligo-IgG bands is that between pH 7 and pH 11 there are normally no other proteins beside the IgGs.

2D-Elpho of Serum

The picture on the left side shows a 2D-separation of serum proteins.
Beside of the IgGs only 3 other proteins can be found between the pH-intervall of interest (6-11):

1. The hemolysate proteins (hemoglobins ) at pH 7-7.2 (16 000)
(It is important to identify the Hemolysate-proteins in a general visualization procedure: A special PAG-gel pH 6-11 has small slots at the edges to pipet diluted hemolytate onto the gel. Hemolysate proteins in the serum or the CSF-lanes can now be tagged)
2. The Prostaglandin D Synth. (ß-trace) at pH 6.8 (25 000)
3. The Cystatin C (g-trace) at pH 11 (35 000),
(This protein can only be found in th CSF and is located outside of the IgG region. No mismatch analysis should be possible), a degradation product is at pH 8 (30 000).

These are the reasons why:
If a general visualization is chosen the amount of bands in the CSF should be more than 2-3 to be typed as positiv.

Gel: IEFGel pH 6-11 24S, visualization: general silver-staining
Samples: E. Dubois, AG
Dr. Rebecca Schüle Dr. Markus Krumbholz, Zentrum für Neurologie Tübingen (Germany)
(*) = MS positive, on the right side Bio-Rad’s Liquichek Spinal Fluid (K)

Mai 30, 2018

GE's Autostainer

Silver staining can be automated with GE’ s Autostainer

IEFGel 6-11 24S - silver-stained