Anti acetyl histone h3 lys9 antibody

MPCCs were fixed with either ice-cold methanol for 10 minutes at 4°C or 4% paraformaldehyde (PFA) for 20 minutes at room temperature. PFA-fixed samples were permeabilized with 0.1% TritonX for 10 minutes at room temperature. Wells were washed twice with PBS, blocked with 2% bovine serum albumin (BSA) in phosphate-buffered saline (PBS), and incubated with primary antibodies for 1 hour at room temperature. Samples were washed with PBS then incubated with Alexa 546-conjugated secondary goat-anti-mouse (Invitrogen) and Alexa 647-conjugated goat anti-rabbit (Invitrogen) for 1 hour at room temperature. Samples were washed with PBS, counterstained with the DNA dye Hoechst 33258 (Invitrogen; 1:5,000), and kept in Aquamount (Lerner Laboratories). Images were captured on a Nikon Eclipse Ti fluorescence microscope or a Nikon 1AR Ultra-Fast confocal microscope. Areas of the developing liver stage parasites were measured using ImageJ and used to calculate the corresponding diameter. Liver stage P. vivax parasites were detected using rabbit polyclonal antibodies against UIS4, BIP, MIF, HSP60, HSP70, MSP1 and mouse monoclonal antibodies against UIS4 and ACP. The nuclei of the parasites were visualized using a rabbit polyclonal anti-acetyl-Histone H3 (Lys9) antibody (Millipore).