Immunofluorescent analysis of ASH2L (green) in HeLa cells (65% confluency). The cells were fixed with 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 1% Triton X-100 for 30 minutes at 37 C, and blocked with 10% Normal Goat Serum in PBS with 0.1% Triton X-100 for 1 hour at room temperature. Cells were stained overnight with the ASH2L mouse monoclonal antibody (Product # 14-5069-82) at 4 C at a concentration of 1.25 µg/mL in blocking buffer followed by a Goat-anti Rat IgG Secondary Antibody, Alexa Fluor 488 (Product # A-11006) at a dilution of 4 µg/mL in blocking buffer for 1 hour at room temperature. Nuclei (blue) were stained with DAPI in Fluoromount G with DAPI (Product # 00-4959-52). Images were taken on a Zeiss Axio Observer.Z1 microscope at 20 x magnification with an Orca-Flash4.0. camera from Hamamatsu. The upper panel shows cells stained with clone AS 4C5. No non-specific staining was observed with rat IgG2a isotype control (lower panel).