General Silver

Elektrophorese-Technik

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Visualization: General Silver Staining

Dez 17, 2008


	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.

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: FocusGel 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: FocusGel 6-11 24S Visualization: General silver-staining Samples: Prof.Reiber, IBA-course, Göttingen 2004, S=Serum, 1-12 CSF, sample #7: Germany Inter lab test 2004 (Courtesy of Dr.Assert, Klinikum Essen)




	Silver staining can be automated with GE" s Autostainer