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Western Blot Forum Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


Western Blot Problem Band Issues

Western Blot Problem Band Issues - Western Blot Forum

Western Blot Problem Band Issues - 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 04-07-2011, 12:12 PM
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Default Western Blot Problem Band Issues



Hello everyone
I am facing a strange kind of problem, generally speaking, we use to use secondary antibodies which are made against the same organism as that for the primary ab, eg- if the primary ab origin is goat we use antigoat 2 ab. This goes fine with me also till... the mouse origin comes. The problem is, when i use goat or rabbit 1 ab, and then anti goat or antirabbit 2 ab, i get the bands, but whenever i use mouse 1 ab and then antimouse 2ab, i dont get anything, i tried this for 4-5 times with diff abs, but always same result, except that for B-ACTIN.
More strange this is that, once i use b-act (mouse origin) (1 and 2 ab both mouse) and after that i streap the blot with striping soln, and i reuse the blot with other mouse origin ab, i get results. But everytime i feel that it is the bact which is coming everytime.
Please suggest me something to solve this problem.
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