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Visualization: Immuno-Staining

Oktober 18, 2021

EDC’s new BlotGels can be used to perform a electro-blot because BlotGels can easily be removed from their support film. See EDC’s new manual.
The semidry blot method is recommended.

After the IEF the support film of the BlotGels should be removed first.
To do this first a dry nitrocellulose membrane 0.45 um (GVS North America, NitroPure, supported, #1212590) is fixed to the lab bench and the gel is placed on it, gel-facing down. The support film is now rolled away and the stack is taken to the blot apparatus. BlotGel IEF-manual.

Then the Immuno-Staining with an Anti-human IgG Antibody (HRP-conjugated, DAKO P0449) is performed on the membrane.

Advantage: A well introduced technique, IGG-selective staining.
Disadvantage: A additional technique (the blotting) has to be perfromed

fixing the membrane

fixing the blot-membrane on the bench

laying on the gel

the gel is placed on the membrane

semidry blotting stack

the semidry blotting stack

rolling away the support

the support film is rolled away

pH 6-11
Immuno Stain on membrane

Gel: BlotGel NF pH 6-11
Visualization: Immuno-staining after contact-blotting
(Courtesy of .A.W. Teelken UMCG, Neurologic Klinic, Groningen)

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