Hi everyone!
I had some trouble in performing a good staining for aurora B kinase on rat cells. I normally did:
-fixing of cells with PFA 4%
-permeabilization with Triton 0.5%
-blocking with BSA 3%
-overnight ab 1:300 in 1.5% BSA with aurora B antibody abcam ab2254
-secondary ab the day later.
What I found is a super bright not specific signal on all the cells (control with no primary antibody is clean).
Can anyone suggest me a better protocol or an antibody that works fine for Immunofluorescence on rat cells ??