HeLa cells were stained with H3pan polyclonal antibody (Cat # PAB14048) and with DAPI. Cells were formaldehyde fixated, permeabilized with sodium citrate and Triton X-100 and blocked with PBS containing 2.5% BSA. (A) Cells were immunofluorescently labelled with the H3 antibody (diluted 1 : 500 and incubated for 1 hour at room temperature) followed by goat anti-rabbit antibody conjugated to DyLight 488. (B) Staining of the nuclei with DAPI, which specifically labels DNA. Both, antibody and DAPI staining are restricted to the nucleus. The antibody is directed against the unmodified form of H3. Note the uniform nuclear distribution of H3 which is a basic component of the nucleosome.
ELISA was performed using a serial dilution of H3pan polyclonal antibody (Cat # PAB14048). By plotting the absorbance against the antibody dilution, the titer of the crude serum was estimated to be 1 : 500.