Human phospho-p53 (S15) Antibody

R&D Systems | Catalog # AF1043

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Immunocytochemistry, Immunoprecipitation

Cited:

Immunohistochemistry, Western Blot, Immunoprecipitation, Westen Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all p53 Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing human p53 S15 site

Specificity

Detects human, monkey, and rat p53 when phosphorylated at S15 and mouse p53 when phosphorylated at S18.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human phospho-p53 (S15) Antibody

Detection of Human Phospho-p53 (S15) antibody by Western Blot.

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) antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

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) antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P).

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) antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P).

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

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

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

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

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.

Applications for Human phospho-p53 (S15) Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line treated with UV light

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human breast cancer tissue subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)

Immunoprecipitation

1 µg/500 µg cell lysate
Sample: CEM human T-lymphoblastoid cell line exposed to UV-C and MCF‑7 human breast cancer cell line treated with camptothecin (CPT), see our available Western blot detection antibodies

Western Blot

0.2 µg/mL
Sample: CEM human T-lymphoblastoid cell line exposed to UV-C

Reviewed Applications

Read 3 reviews rated 4.3 using AF1043 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: p53

The p53 tumor suppressor protein is a multi-functional transcription factor that regulates proliferation, cell cycle checkpoints, and apoptosis. The importance of p53 is underscored by its mutation in over 50% of human cancers. Human p53 is a 393 amino acid (aa) nuclear/cytoplasmic protein that contains an N-terminal activation domain, a specific DNA binding domain, and a C-terminal domain that mediates tetramer formation and regulates DNA binding. P53 is phosphorylated on Ser15 in response to irradiation, and this phosphorylation increases recruitment of the coactivator CREB-binding protein/p300. The ATM or ATR kinases can phosphorylate p53 at serine 15 (S15), which leads to cell cycle arrest. Serine 15 phosphorylation leads to p53 stabilization and enhances transactivation of p53 target genes.

Alternate Names

BCC7, LFS1, TP53, TRP53

Entrez Gene IDs

7157 (Human); 22059 (Mouse); 24842 (Rat)

Gene Symbol

TP53

Additional p53 Products

Product Documents for Human phospho-p53 (S15) Antibody

Certificate of Analysis

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Product Specific Notices for Human phospho-p53 (S15) Antibody

For research use only

Citations for Human phospho-p53 (S15) Antibody

Customer Reviews for Human phospho-p53 (S15) Antibody (3)

4.3 out of 5
3 Customer Ratings
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4 Stars
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  • Human Phospho-p53 (S15) Antibody
    Name: Gabriele Strusi
    Application: Western Blot
    Sample Tested: HepG2 human hepatocellular carcinoma cell line
    Species: Human
    Verified Customer | Posted 03/02/2023
    Suggested concentration was to low, increased to get better results
    Human phospho-p53 (S15) Antibody AF1043
  • Human Phospho-p53 (S15) Antibody
    Name: Qing Xu
    Application: Western Blot
    Sample Tested: Cancer Cells
    Species: Human
    Verified Customer | Posted 07/11/2018
    PC3 cells were treated with UV light for the indicated times. Total cell lysates were subjected to western blot. PVDF membrane were probed with 1mm/ml Human Phospho-p53 (S15) Antibody (AF1043). A specific band was detected for p-P53 at approximately 53kDa. This experiment was conducted under reducing conditions
    Human phospho-p53 (S15) Antibody AF1043
  • Human Phospho-p53 (S15) Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: MDA-MB-231 human breast cancer cell line and T47D human breast cancer cell line
    Species: Human
    Verified Customer | Posted 08/16/2017

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