EDC Impressum & DSE

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Mai 30, 2018

General:
I
n native and in SDS-electrophoresis the Immunoglobulin G fraction (IgG) shows only 1 slightly broadened uniform band. Different oligoclonal IGG-fractions can only be separated from each other when the isoelectric focusing (IEF) is chosen as separation technique. In the isoelectric focusing the proteins were separated according to their charges along a pH-gradient. So very small differences in the isoelectric points of the proteins show different running distances.
The cerebrospinal fluid (CSF) is an immuno-privileged space. Normally only a few T-Lymphocytes migrate through the blood-brain barrier. In the case of a disease the amount of Lymphocytes increases in the CSF.
So both, the serum and the CSF per patient has to be determined. If oligoclonal IgG fractions occur in the CSF and not in the cooresponding serum, specific metabolic processes are going on.
Results:
Please see the different visualization procedures below.
Standards: Bio-Rad Liquichek™ Spinal Fluid Control, Level 1 #751, Level 2 #752

 horizontal polyacrylamide IEF-gel

FocusGel24S

Special PAG-gel 6-11 with slots:
S
pecial gels for Oligo-IgGs with preformed slots for 24 x 25ul (24S) res. 40 x 12ul (40S) sample volume.
The pH-gradient is optimized and spreaded in the region between 6 and 11. Please compare with competitor #3
FocusGels are washed, dried and rehydrated gels, ready to use for the isoelectric focusing.
These gels are not toxic! The ampholine-cocktail forming the pH-gradient in the electric field was never in contact with the polymerization chemicals: No destroying and no aging of the ampholines occurs as low resolution and thick brown stripes after the staining.
Highest resolution of all bands, see figure below. This is not possible with Agarose-based IEF gels, see the gels of competitors #1 and #2.

The Visualizations:

[General Silver]
[Immunfluoresc.]
[Immunofixation]
[Immunostain]

The interpretation:

The Competition:

[Interpretation]
[Competition]