Flow cytometric analysis of Phospho-Histone H3.3 (S31) expression in HepG2 cells using Phospho-Histone H3.3 (S31) antibody (Cat#61418, 1:2,000). Green, isotype control; red, Phospho-Histone H3.3 (S31).

Immunocytochemical staining of HepG2 cells with anti-Phospho-Histone H3.3 (S31) antibody (Cat#61418, 1:1,000). Nuclei were stained blue with DAPI; Phospho-Histone H3.3 (S31) was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Scale bar, 20 μm.

Western blotting analysis using anti-phospho-Histone H3.3 (S31) antibody (Cat#61418). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-phospho-Histone H3.3 (S31) antibody (Cat#61418, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).

Western blotting analysis using anti-phospho-histone H3.3 (S31) antibody (Cat#61418). Phospho-histone H3.3 (S31) expression in wild type (WT) and H3-3A knockdown (KD) HSHC cells with 5 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with anti-phospho-histone H3.3 (S31) antibody (Cat#61418, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).

Immunohistochemistry was performed on paraffin-embedded human sigmoid colon carcinoma using anti-phospho-histone H3.3 (S31) antibody (Cat#61418, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar: 25 μm.