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Convex shape of sample bands on SDS PAGE gel

Convex shape of sample bands on SDS PAGE gel - Western Blot Forum

Convex shape of sample bands on SDS PAGE gel - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 04-20-2007, 09:43 AM
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Default Convex shape of sample bands on SDS PAGE gel



Dear all
sometimes i get convex shape of sample bands while running SDS PAGE.
Is it due to change in pH of buffers used in SDS PAGE gel prepn?

Thanks
ssacket
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Old 05-28-2007, 04:14 PM
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Default Re: Convex shape of sample bands on SDS PAGE gel

Hi!
Sometimes convex shape results from gel overloading (usually I don't load more than 50 ug of protein for each lane), or you should run your gel slower than 100 V.
I never had problems with buffers.....but I store them always at +4°C.
Good luck!
m.
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Old 10-30-2007, 11:59 PM
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Default Re: Convex shape of sample bands on SDS PAGE gel

I had the same issue (convex bands) in my western blot.

I did load 50ug-60ug protein, and only 2 of 8 lanes resulted in convex shape.
I ran it at 100V.

Overloading could have been the reason, but it is unclear since only 2 of the 8 bands had convex shape.

Any other suggestions anyone?

Thanks!!
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Old 10-31-2007, 06:33 PM
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Default Re: Convex shape of sample bands on SDS PAGE gel

Hello,
also you should make sure you load loading buffer in each lane including the ladder ( protein marker) as salt concentrations can also affect the sharpness and flatness of your bands.

Cheers!
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Old 12-04-2007, 01:09 AM
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Default Re: Convex shape of sample bands on SDS PAGE gel

That usually happens when you run your gels at too high of a voltage. It can also be due to your migration buffer so make sure your buffer is well mixed and homogenous. In my case, it usually evens out before it reaches the bottom so it doesn't cause any problems.
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