Caveolin-3 western bands??

[agador]

Hi there,

I'm getting multiple bands in my western blots for caveolin-3. The manufacturer states bands should appear in the 20-25 kDa... and they certainly do; the problem being there is multiple signals in this range. Does anyone have experience with western blots for caveolin-3 or any of the other caveolins. Although these signals may be non-specific bands I am also speculating that these other bands may be caveolin complexes with other proteins or cholesterol. Apologies for posting in the wrong area. I guess the western's forum would have been better but I am also interested in structure/function reason(s) why caveolin migration would be so variable.

Thanks in advance
Anthony

[admin]

Hello there welcome to the forums,

are there any phosphorylated or modified PTMs of caveolin-3? that may be the issue.

Also try incubating blotto milk 2-5% during your primary and secondary antibody incubations that may clear up the issue.

When you expose quickly do you still see these bands? Also did your film move at all, that could be a simple problem that causes multiple bands.

Please let us know also posting a gel would help us problem solve it.

cheers!