hey guys I am using strep II tag for detection of an heterologously expressed protein. On western analysis I am getting exact band even in non transgenic wild type strain. I have checked my wild type strains thoroughly, to ensure any kind of cross contamination. Even PCR amplification for the transgenes in wild type strain is negative. Do anyone have any troubleshoot idea or an possible explanation for this situation?
Could it be possible that the transgenic samples were overflown (spilled) into the non-transgenic sample lane(s) while you were running a gel for Western blotting??
No i dont think so, in such case intensity of the fake band would be very less
If it is polyclonal, you may have a specific binding to unfortunately an approximately same sized protein. You could try to subtract that fraction from your antibodies using WT protein extract, or try a different antibody.
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