2dtrouble

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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		2D-Electrophoresis Troubles

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


	April 03, 2025


	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!


	[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


	EDC Electrophoresis Development & Consulting, Vor dem Kreuzberg 17, 72070 Tübingen (Germany)