Good Antibody Against Phospho H2A.X

Used this antibody for immunofluorescence imaging of cells treated with DNA damaging agents. Antibody works well, but there is a slight hue on the DNA if you don't pre-extract. The monoclonal version of this antibody is much more specific. This is the protocol used: Cells were incubated with Microtubule stabilizing buffer (MTSB) for 1 minute Permeablized with MTSB + 0.5% Triton X for 2 minutesIncubated with MTSB for 1 minuteCells 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 PBS 10 minutes in PBS+3% BSA Antibody was incubated in PBS+1% BSA and coverslips incubated for 1 hour 2 - 5 minute washes in PBS Secondary antibodies from Jackson Laboratories (anti-rabbit) was incubated in PBS+1% BSA and coverslips2 - 5 minute washes in PBS Mounted coverslips using ProLong Gold