Western blot bands look like "M" - (Mar/17/2006 )

I have read some posts on here that suggest it could be a problem with the gel??? But why are some lanes misshaped and others are perfect?

Thanks

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perhaps uneven polymerization? (similar effect would be seen in an agarose gel if there were little lumps that didn't melt properly, right?)

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i've seen similar effect when "skins" are left in the sample wells by the combs (some combs fit unevenly). if that is the cause then you might be able to defeat it by clamping the plates on the teeth of the comb or by scraping any skin out of the well prior to sample introduction.

this is assuming that all your samples are the same. differing conditions (salt, additives, etc) of the sample could also cause strange shaped bands.

by the way, is that a gel or a blot in the picture. if it's a blot then those funny bands could be caused by bubbles between the gel and membrane.

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Are the same lanes consistantly warped from gel to gel?

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My first answer was going to be no, but when I went back and looked at the last 2 blots the same 4 lanes in both were warped. Could this be more than a coincidence?

However, previously I have had all of the lanes or just one lane that is warped.

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can also be caused by warped plates

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If the same lanes are consistantly warped, you can pretty much rule out the more random stuff, like flaws in polymerization, etc.

In those instances, the first place I'd look would be the little wire that the bubbles come off of... if you have bits of gel or dirt clinging to it you can create an uneven electrical gradient....

Next, I'd try running and pouring my gel elsewhere on the bench (you could have an unlevel bench.)

That's assuming it's in the page step...

Next, get new sponges and filterpaper for your transfer sandwich... see if that makes a difference.

If none of those work.... worst case scenario is you need new equipment....

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