western blot - (Aug/10/2007 )

Why is there a need to sandwich the gel with 4 or more pieces of filter paper?
Thank you.

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wet tranfert : to ensure the pressure will be homogenized on the gel
semidry transfert : same as wet + not drying of the gel and the membrane during the transfert

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it is only necessary for semi-dry blotting: paper work as depot for transfer buffer;

in tank blotting, you need only 1 piece of paper per side

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In semi-dry, you can put 4-6 filter papes, but do not put more than 6 filter papers as protein in gel will not transfer to the membrane...may be too thick for the current to pass through...

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You could use only one filter paper on each side.

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Hi,
I am a bit confused about these terms, semi-dry, wet transfer and tank blotting. I lay sponges, filter papers, gel and membrane on a cassette and put it in a tank and pour transfer buffer into the tank. I guess this should be called tank blotting. Am I right? Is this considered also wet transfer?
Thank you.

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Yes, tank blotting and wet transfer mean the same type, filling a tank with transfer buffer. The other option is semidry, carried out in a different apparatus (semidry trasnfer units) using less amount of transfer buffer and usually completed in less time.

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