EDC Impressum & DSE

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Dr. Hanspeter Schickle
Wolfgang Gstrein

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IEF, native: Salt in Samples

September 21, 2020

Symptom: A sample lane is spreaded lateral displacing their neighbour lanes.
Cause: When samples hold too much salt they are
a) accelerated lateral towards the gel without salt-loading, lateral sample spreading
b) SDS: unsharp band
1.   Tolerate this.
2.   IEF, native electrophoresis: Desalt sample using small columns like NAP-5 (GE 17-0853-01)

IEF and native sample-desalting procedure:

[salt in the samples]

3. SDS: Precipitate the proteins with 20% TCA. Centrifuge samples. Wash 1 x with dist.water. Centrifuge. Resolubilize with sample buffer.
Additional advantage beside the desalting effect: sample concentrations can be increased easily!

....a salty IEF-sample is spreading lateral

....Urines (11-6), after TCA-precipitation (and concentrating)