Hello guys:
Just a very silly and practical question: how to get rid of the bubble during western blot transfering? I know several tricks, but none of them worked perfectly. So if the bubble locate in the bands coincidently, which means another confirmation or validation, it is totally time-wasting.
Any smart people want to share your precious tricks which guarantee no bubble in the blot?
Many thanks in advance.
Shiqiu Xiong
roll a 15ml falcon tube (without the lid) on your sandwich while holding it in place by putting the tip of your finger on the middle of one of the extremities.
Thanks Henri:
We have specifically designed roller, it seems not perfect for me.
Regards
It's hard to say without looking at how you assemble the transfer cassette. I've never used rollers, I just assemble each successive piece of the sandwich (filter papers, membrane, gel, then filter papers again) by ensuring that a film of buffer is covering the previous piece and then bending the next one at the center, touching the previous piece, then lowering each side slowly to displace any existing bubbles.
Also, I prewet membranes, papers, etc. by first floating them on water for a minute and then immersing them for another minute. Same with foam pads.
Hope it helps, I'm not sure I'm making much sense here.
Dear Alejandro:
Thanks for your sharing. I got your points. Soaking the film and touching gel from the centre may be helpful. I will try and compare.
Thanks again.
A good idea is to prepare all the sandwich sumerged in a container filled with tbs buffer or transfer buffer
Dear Shiqiu
My tricks is that after rolling the roller, I slowly lift the filter paper and look for no bubbles. If there are bubbles, I remove them manually with a wet filter paper edge, pressing directly on the PVDF membrane. Then I adjust the sandwich and roll the roller again.
Thanks Julia:
That's I normally try, it can get rid of big bubbles, but didnt work for small one. Maybe I am not careful enough. Thanks for sharin.
Best wishes.
Dear Emilio:
I just tried to prepare film and gel in transfer buffer, it is easy to manipulate and no bubbles at all. The best!
Thanks for your idea.
Shiqiu
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