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Use of colloidal coomassie stained to western blotting?? - (Jul/12/2011 )

Hi All,
Can one use a colloidal coomassie stained SDS-PAGE gel for the western blotting?
I mean, once the gel is coomassie stained, then can we destain it with destaining solution and use the same for the western blottin?
Have anyone tried it.
Please give your suggestions about it.
Thank you

-Myco Rax-

the coomassie stain may interfere with the western, especially if you develop with color deposition.

the coomassie will, at least partially, come off the protein when transferred but will bind to the membrane.

-mdfenko-

mdfenko on Fri Jul 15 16:38:11 2011 said:


the coomassie stain may interfere with the western, especially if you develop with color deposition.

the coomassie will, at least partially, come off the protein when transferred but will bind to the membrane.

Hi,
What if I completely destain it?
Thank you

-Myco Rax-

if u just wanna identify ur protein of interest then u can go ahead.. hoping that the antibody binding site is secure!!!

-Prep!-

Myco Rax on Mon Jul 18 07:40:55 2011 said:


What if I completely destain it?

can you? i think that may be very difficult.

you can transfer the stained protein (it will destain some during transfer). it's just that it will interfere with seeing color deposited on the membrane. if you are looking for fluorescence or chemiluminescence then you should have no problem.

-mdfenko-

For me, the amount of protein necessary to see by coomassie is way higher than that needed to see by western blot.

-kfunk106-