Good Antibody Against Bub1

Antibody was used to mark prometaphase cells. Cells were fixed and stained with ACA antibody (Anti-centromere antibody) and DNA. Antibody can be used at both room temperature for 1 hour or overnight at 4C. Antibody seems to be very specific and overall worked really well. Here is the protocol that I used: Cells were fixed in 3.5% paraformaldehyde for 15 minutes at room temperature.3 minutes in ice cold methanol. 5 minutes in PBS+0.5% Triton X5 minutes in PBS10 minutes in PBS+3%BSAAntibody was incubated in PBS+1%BSA and coverslips incubated for 1 hour 2 - 5 minute washes in PBSSecondary antibodies from Jackson Laboratories (anti-rabbit) was incubated in PBS+1%BSA and coverslips2 - 5 minute washes in PBS Mounted coverslips using ProLong Gold.