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Strange bubble seen on Sypro blot staining

Strange bubble seen on Sypro blot staining - Western Blot Forum

Strange bubble seen on Sypro blot staining - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 02-23-2010, 11:45 AM
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Default Strange bubble seen on Sypro blot staining



Hello everyone,

since a couple of weeks we have problems with the detection of our western blots (bands are deformed). We did a Coomassie staining on the gel to see if the problem was due to the running which wasn't the case. We also did Syprostaining on several blots to see if there are problems with the transfer. When do so, there are strange bubbles visible. These bubbles are always on different places. But when detecting proteins, which are in the range of the bubble, the deformation of the bands is visible.
I've put some pictures with this thread just to make my story more clear.
Please help me find the problem of our western blot. Every suggestion is welcome!

thanks in advance!
Attached Images
File Type: jpg detection with NBTBCIP kit on same gel.jpg (24.5 KB, 15 views)
File Type: jpg syprostaining.JPG (17.9 KB, 12 views)
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Old 02-26-2010, 11:43 PM
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Default Re: Strange bubble seen on Sypro blot staining




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Old 02-26-2010, 11:45 PM
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Default Re: Strange bubble seen on Sypro blot staining

Hi there,
looks like a transfer/membrane/gel issue all around.

I would carefully re-do your buffers, pH them. Make sure when making your gels you mix well. Separate at constant current.

Also for transferring keep out any bubbles by rolling.

For membranes, make sure you keep them clean and don't get them dirty making sure the whole membrane is covered by solutions when shaking.
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