Human Phospho-p53 (S15) Antibody

Detection of Human Phospho-p53 (S15) by Western Blot. Western blot shows lysates of CEM human T-lymphoblastoid cell line untreated or exposed to 100 J/m²UV-C for the indicated time. PVDF membrane was probed with 0.2 µg/mL Human Phospho-p53 (S15) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1043) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Phospho-p53 (S15) was detected at approximately 53 kDa (as indicated). The phospho-specificity of this antibody was supported by decreased labeling following treatment with 600 U lambda-phosphatase (lambda-PPase) for 1 hour. For additional reference, duplicate lysates were probed with 1:5000 dilution Human/Mouse/Rat p53 HRP-conjugated Antigen Affinity-purified Polyclonal Antibody (lower panel, Catalog # HAF1355). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Phospho-p53 (S15) in HeLa Human Cell Line. p53 phosphorylated at S15 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with UV light (left panel) or untreated (right panel) using Rabbit Anti-Human Phospho-p53 (S15) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1043) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. Cells were co-stained using Rat Anti-Tubulin (Catalog # NB600-506, Novus Biologicals) and NorthernLights™ 493-conjugated Anti-Rat IgG Secondary Antibody (green; Catalog # NL015). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Phospho-p53 (S15) in Human Breast Cancer Tissue. p53 phosphorylated at S15 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Human Phospho-p53 (S15) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1043) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of immersion fixed paraffin-embedded Tissue Sections.

Phospho-p53 (S15) in Human Breast Cancer Tissue. p53 phosphorylated at S15 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Human Phospho-p53 (S15) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1043) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of immersion fixed paraffin-embedded Tissue Sections.

Detection of Phospho-p53 (S15) by Western Blot The increase of HDAC9 was associated with bone and fat imbalance in bone aging. a Micro-CT analyses of bone mass of trabecular and cortical bone thickness in the femora of 2-month-old (young) and 16-month-old (aged) mice. Bone mineral density (BMD), trabecular bone volume (BV/TV), and cortical bone thickness (Ct.Th) were performed. Scale bar = 1 mm. b–d Immunofluorescent staining of OCN (b), PPAR-gamma (c), and TRAP (d) was performed in the bone marrow from young and aged mice, and the positive signals were quantitatively analyzed. Scale bar = 50 μm. e Expressions of the senescence-related proteins, p53 and p-p53, in the bone marrow from young and aged mice were examined by western blotting. f Expressions of the senescence-related proteins, p53 and p-p53, in BMMSCs from young and aged mice were examined by western blotting. g Alizarin Red staining was performed, and quantification of mineralized nodules was analyzed in young and aged BMMSCs. h Oil Red O staining was performed, and quantification of lipid droplet positive ratio areas was analyzed in young and aged BMMSCs. Scale bars = 100 μm. i, j Expression of HDAC9 (i) and acetylation sites of H3K, including H3K9, H3K18, and H4K16 (j), were examined by western blotting. The data are presented as the means ± SD of each independent experiment performed in triplicate. *P < 0.05, **P < 0.01, ***P < 0.001, unpaired two-tailed Student’s t test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32620134), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Phospho-p53 (S15) by Western Blot Downregulation of HDAC9 rescued lineage differentiation imbalance and ameliorated senescence in aged BMMSCs. a Alizarin Red staining was performed, and osteogenesis-related proteins were detected by western blotting in aged BMMSCs transfected with HDAC9 siRNA. b Oil Red O staining was performed, and adipogenesis-related proteins were detected by western blotting in aged BMMSCs transfected with HDAC9 siRNA. c Alizarin Red staining was performed, and osteogenic-related proteins were detected by western blotting in young BMMSCs and young BMMSCs transfected with HDAC9 siRNA. d Oil Red O staining was performed, and adipogenic-related proteins were detected by western blotting in young BMMSCs and young BMMSCs transfected with HDAC9 siRNA. e, f Expressions of the senescence-related proteins p53 and p-p53 in BMMSCs cultured in vitro from aged mice (e) and young mice (f) were examined by western blotting. The data are presented as the means ± SD of each independent experiment performed in triplicate. *P < 0.05, **P < 0.01, ***P < 0.001, one-way analysis of variance (ANOVA) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32620134), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Phospho-p53 (S15) by Western Blot Downregulation of HDAC9 rescued lineage differentiation imbalance and ameliorated senescence in aged BMMSCs. a Alizarin Red staining was performed, and osteogenesis-related proteins were detected by western blotting in aged BMMSCs transfected with HDAC9 siRNA. b Oil Red O staining was performed, and adipogenesis-related proteins were detected by western blotting in aged BMMSCs transfected with HDAC9 siRNA. c Alizarin Red staining was performed, and osteogenic-related proteins were detected by western blotting in young BMMSCs and young BMMSCs transfected with HDAC9 siRNA. d Oil Red O staining was performed, and adipogenic-related proteins were detected by western blotting in young BMMSCs and young BMMSCs transfected with HDAC9 siRNA. e, f Expressions of the senescence-related proteins p53 and p-p53 in BMMSCs cultured in vitro from aged mice (e) and young mice (f) were examined by western blotting. The data are presented as the means ± SD of each independent experiment performed in triplicate. *P < 0.05, **P < 0.01, ***P < 0.001, one-way analysis of variance (ANOVA) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32620134), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Phospho-p53 (S15) by Western Blot The increase of HDAC9 was associated with bone and fat imbalance in bone aging. a Micro-CT analyses of bone mass of trabecular and cortical bone thickness in the femora of 2-month-old (young) and 16-month-old (aged) mice. Bone mineral density (BMD), trabecular bone volume (BV/TV), and cortical bone thickness (Ct.Th) were performed. Scale bar = 1 mm. b–d Immunofluorescent staining of OCN (b), PPAR-gamma (c), and TRAP (d) was performed in the bone marrow from young and aged mice, and the positive signals were quantitatively analyzed. Scale bar = 50 μm. e Expressions of the senescence-related proteins, p53 and p-p53, in the bone marrow from young and aged mice were examined by western blotting. f Expressions of the senescence-related proteins, p53 and p-p53, in BMMSCs from young and aged mice were examined by western blotting. g Alizarin Red staining was performed, and quantification of mineralized nodules was analyzed in young and aged BMMSCs. h Oil Red O staining was performed, and quantification of lipid droplet positive ratio areas was analyzed in young and aged BMMSCs. Scale bars = 100 μm. i, j Expression of HDAC9 (i) and acetylation sites of H3K, including H3K9, H3K18, and H4K16 (j), were examined by western blotting. The data are presented as the means ± SD of each independent experiment performed in triplicate. *P < 0.05, **P < 0.01, ***P < 0.001, unpaired two-tailed Student’s t test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32620134), licensed under a CC-BY license. Not internally tested by R&D Systems.