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Vertical streakings on 2-D gel (probably glycoprotein)

Vertical streakings on 2-D gel (probably glycoprotein) - Proteomics Forum

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Old 10-26-2006, 09:05 AM
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Default Vertical streakings on 2-D gel (probably glycoprotein)



I have got very strong vertical streakings on my 2-D gel, sb. in our lab suggested that the spots involved may be glycoproteins.
The result is consistently reproducible.

Have anybody ever dealed with this kind of problem?
Or could it be other problem?

Thanx
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Old 10-27-2006, 12:26 AM
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Default Re: Vertical streakings on 2-D gel (probably glycoprotein)

Hey there Shepherdcat!,

I have some experience with this. Yes it may probably be a glycoprotein. Did you try any other buffers and/or dialysis of the samples before running it?

If it is a glycoprotein and this is causing it, you could easily treat with a deglycosylase. If that doesnt solve your 2-D problem, I would start looking at other things.

Cheers + welcome here!

Darkwng
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Old 10-27-2006, 04:58 AM
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Default Re: Vertical streakings on 2-D gel (probably glycoprotein)

Darkwng

Thank you for the reply.

No, I haven't try other buffer or dialysis. Other spots on the gel are well-focused except for those problematic ones. So I think it should be sth related to the several specific protein themselves.

I think I will try deglycosylase first but I wonder if it will cause some unwanted effect.

Shepherdcat
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Old 12-07-2006, 07:55 AM
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Default Re: Vertical streakings on 2-D gel (probably glycoprotein)

Thanks! I have the same problems.
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Old 12-07-2006, 09:13 AM
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Default Re: Vertical streakings on 2-D gel (probably glycoprotein)

Hey there Xiaoxiangwen,

any more ideas? do you think its glycosylation that is causing your vertical streaks? I also read a 2-D gel electrophoresis manual.

Vertical Streaking can also be caused by:

Insufficient Equilibration (solution is to increase equilibration time)
Second Dimension Buffer is not properly made (prepare fresh 2-Dimension buffer)
Vertical streaking on 2-D can also be caused by insufficient SDS in electrophoresis buffer (increase to 0.1% SDS)

Last edited by admin; 12-07-2006 at 09:16 AM.
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