EDC Impressum & DSE

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Horizontal Striking
Streaking of spots in the IEF-direction

Spots are striking in the direction: sample application point ---> focusing point
TS-HorizontalStrikingPlusKlein

April 28, 2023

Remedy:
1. General:
Proteins are “dying” during migration from the starting point to the focusing point.
The volt-level was raised to early and too quick! Strips get too hot.
Do not raise the volt-level over 1000 Volt during the first 12 hours.

In our lab only this running condition below run properly, all variations of this running condition failed:

24 cm IPG-strip, cup-loading, total time: 16 h (start 5 pm - stop 9 am)

Volts

uA/strip

µA-start/strip

time

temp

150

80

~ 25 uA

3 h

20°C

300

(80)

~ 50 uA

3 h

20°C

300 --->1000

(80)

~ 30 uA

6 h

20°C

1000 --->10.000

(80)

~ 20 uA

1 h

20°C

10.000

(80)

~ 15 uA

3 h

20°C

---> = Volt gradient

2. Gradients including basic pH-values:
When pH-ranges over pH 7 are run use Bishydroxyethyldisulfide without other reducing agents
Point 1 (above) is also valide!

(only for cup loading)

 S 40750 Vh, 16 h

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