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.