EDC Impressum & DSE

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

Datenschutzerklärung >>

2D-Electrophoresis Troubles

Not many things can be done wrong in 2D electrophoresis!
Most frequently done mistakes:

Mai 30, 2018

Gel is leaving its support film:
Do not use staining recipes with alcohol concentrations higher than 40%.
The gel will shrink or swell to much so the forces between the film and the gel get to high!

Troubles-MeOH

[1D uncomplete]
[heat/Watt-ver]
[heat/Watt-hor]
[air bubble-IPG]
[forward striking]
[backward striking]
[optimal shutdown]
[blue/black curtain]
[DIGE: doublespots]
[buffer-system mix]
[bad or no contact]
[fingerprint]

The first dimension is uncomplete, horizontal striking

Too much heat in the vertical second dimension

Too much heat in the horizontal second dimension

Air bubble(s) underneath the IPG-strip (vertical)

The spots show forward striking

The spots show backward striking

The spot-pattern is compressed

Blue/black curtain after Coomassie- or silver staining

 Derivating the proteins/peptides changes mol-weights

Ugly border enters the gel / buffer system mix-up

Contact(s) between gel and electrode(s) is (are) bad

finger prints .....

150 ug E.coli, visualization: fluorescence after separation

webCoomassie150ugKlein