IPG problem

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Tegrion
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IPG problem

Hi!

Please, ask for help!

Does anybody know possible reason(s) for boiling of mineral oil above IPG? It occurs not for all lanes in simultaneous analysis of 6 IPGs. I cann't see mechanism of reaction...

We use a standard technique from BioRad. But sometimes we don't use wicks.

Ripal
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Hi, Tegrion

Hi, Tegrion
I don't know your meaning of boiling of mineral and you not use wicks.Why?
Now I also used Bio-Rad, and please give some comments about my results

Tegrion
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Hi, Repal!!

Hi, Repal!!

To a pity, we could not found out reasons for boiling of oil. And working w/o wicks was just an experiment, we tried to truncate loss of  time. Now we come back to almost standart protocol of BioRad.

But with some corrections (experience of our colleagues).

Now we use V=3500 V and around 20 h till 60 000 V*H appr. This is more mild condiiotns for proteins in IPG than 10 000 V.

Ripal
Ripal's picture
Hi,Tegrion

Hi,Tegrion
Can you help me find out some problem reasons of my map?
Thank you!

Tegrion
Tegrion's picture
Our collective try to help

Our collective try to help you. Could you tell what is load of protein, gel percentage, sample buffer?
PLease, see attach, hope something will be useful for you. And for more information looking for appropriate topics on this forum.

Some possible reasons are: 1) too fast casting the gel, not laminar flow in center and edges of the sandwich; 2) protein overloading.