Human Phospho-p53 (S15) Antibody

(3 citations)
(1 Review)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human, monkey, and rat p53 when phosphorylated at S15 and mouse p53 when phosphorylated at S18.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Phosphopeptide containing human p53 S15 site
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.2 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunoprecipitation
    1 µg/500 µg cell lysate
    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
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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/m2 UV-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.
Immunocytochemistry
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.
Immunohistochemistry
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.
Immunohistochemistry
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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
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.
  • Entrez Gene IDs:
    7157 (Human); 22059 (Mouse); 24842 (Rat)
  • Alternate Names:
    Antigen NY-CO-13; BCC7; FLJ92943; LFS1; LFS1TRP53; p53 tumor suppressor; p53; P53cellular tumor antigen p53; Phosphoprotein p53; TP53; transformation-related protein 53; TRP53; tumor protein p53; Tumor suppressor p53
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. Skeletal muscle DNA damage precedes spinal motor neuron DNA damage in a mouse model of Spinal Muscular Atrophy (SMA).
    Authors: Fayzullina, Saniya, Martin, Lee J
    PLoS ONE, 2014;9(3):e93329.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  2. Noninvasive assessment of regulable transferred-p53 gene expression and evaluation of therapeutic response with FDG-PET in tumor model.
    Authors: Aung W, Hasegawa S, Koshikawa-Yano M
    Gene Ther., 2010;17(9):1142-51.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. The p53 oncoprotein is a substrate for tissue transglutaminase kinase activity.
    Authors: Mishra S, Murphy LJ
    Biochem. Biophys. Res. Commun., 2005;339(2):726-30.
    Species: Human
    Sample Type: Recombinant Protein
    Application: WB
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Ctrl AB-105-C 43  
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Ctrl, CyTOF-ready MAB1050  
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Rabbit IgG HRP-conjugated Antibody

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Donkey Anti-Rabbit IgG NorthernLights™ NL557-conjugated Antibody

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Average Rating: 4 (Based on 1 review)

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Summary

ApplicationWestern Blot
Sample TestedMDA-MB-231 human breast cancer cell line,T47D human breast cancer cell line
SpeciesHuman

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