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No SDS in gels - (Jun/25/2010 )

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I've never seen this kind of crackling as in the picture thus i'm guessing.
I,m not sure but looking at the picture i have the feeling that the problem originates from the gel casting or the imager. I don't think the blotting or the probing caused this problem.

-Gerard-

mdfenko on Jul 1 2010, 06:27 AM said:

why do you change blocking agent between antibodies? with what do you block the membrane?

you may want to consider adding normal sheep serum to the block and antibody solutions.



I use BSA with my primary Ab because we re-use our primary antibodies after probing. Sodium Azide is added to it to preserve it for future use.

-Leon_K-

Gerard on Jul 2 2010, 09:59 AM said:

I've never seen this kind of crackling as in the picture thus i'm guessing.
I,m not sure but looking at the picture i have the feeling that the problem originates from the gel casting or the imager. I don't think the blotting or the probing caused this problem.


You're right, after help from a colleague, it seems that i've used the wrong settings to image the blot. Also, i used regular ECL which gave a very weak signal. After using Super ECL, the bands were much clearer.

-Leon_K-
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