Western blotting analysis using phospho-p53 (S33) antibody (Cat#61265). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with phospho-p53 (S33) antibody (Cat#61265, 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 phospho-p53 (S33) antibody (Cat#61265). Phospho-p53 (S33) expression in wild type (WT) and TP53 knockdown (KD) HSHC cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with phospho-p53 (S33) antibody (Cat#61265, 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).
Flow cytometric analysis of Phospho-p53 (S33) expression in HeLa cells using Phospho-p53 (S33) antibody (Cat# 61265, 1:2,000). Green, isotype control; red, Phospho-p53 (S33).
Immunocytochemical staining of Hela cells with Phospho-p53 (S33) antibody (Cat#61265, 1:1,000). Nuclei were stained blue with DAPI;Phospho-p53 (S33) was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain:High. Scale bar, 20 μm.
Immunohistochemistry was performed on paraffin-embedded human sigmoid colon carcinoma using phospho-p53 (S33) antibody (Cat#61265, 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.
