equilibration

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

Datenschutzerklärung >>

Equilibration of the IPG Strip




	IPG-Strip Equilibration Buffer




	Equilibrator-trenches, left side: 12 x 3 ml

April 28, 2023

IPG-Strip Equilibration

Weigh in the Urea and the DTT*, res. the IAA** in 2 test tubes (marked with “DTT” and “IAA”),
then add the 6 ml IPG-Strip Equilibration Buffer. Shake till the Urea has been solved (~10 min).
Then add the solutions in 2 tubes (marked with “DTT” and “IAA”) long enough to take the IPG-strips.

Insert the IPG-strip(s) in the Equilibrator, gel side looking up, then add 3 ml of the first (DTT) equilibrating-solutions and shake horizontally each step for 15 min. Repeat this for the second (IAA) equilibrating solution.
See picture above.
* Dithiothreitol **Iodoacetamide

Step		Urea	Thiourea	DTT*	IAA**	Equilibr. Buffer	Time
1	per strip	2.2 g	915 mg	60 mg	---	6 ml (makes ~7.2 ml)	15 min (shake gently)
2	per strip	2.2 g	915 mg	---	150 mg	6 ml (makes ~7.2 ml)	13 min (shake gently)
3		---	---	---	---	6 ml gel-buffer	2 min (shake gently)


	Normally used: 1 6 M Urea, 2% SDS, 30% glycerol, 50 mM Tris-Cl pH 8.8, 1% DTT, 0.01% Bromophenolblue, 15 min 2 6 M Urea, 2% SDS, 30% glycerol, 50 mM Tris-Cl pH 8.8, 2.5% IAA*, 0.01% Bromophenolblue, 15 min


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