Human/Mouse/Rat p53 Antibody AF1355: R&D Systems

Human/Mouse/Rat p53 Antibody

(6 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat p53.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human p53
    Asp7-Asp393
    Accession # P04637
  • 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
    0.25 µg/mL
    See below
  • Simple Western
    2.5 µg/mL
    See below
  • Immunoprecipitation
    1-2 µg/500 µg cell lysate
    CEM human T-lymphoblastoid cell line, see our available Western blot detection antibodies
  • Chromatin Immunoprecipitation (ChIP)
    5 µg/5 x 106 cells
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Knockout Validated
    p53 is specifically detected in HeLa humancervical epithelial carcinoma parental cell line but is not detectable inp53 knockout HeLa cell line.
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 p53 by Western Blot. Western blot shows lysates of CEM human T‑lymphoblastoid cell line and MCF‑7 human breast cancer cell line were mock-treated (-) or exposed (+) to 10 Gy ionizing radiation (IR) and harvested after 1 hour. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat p53 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1355), followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for p53 at approximately 53 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Chromatin Immunoprecipitation (ChIP)
Detection of p53-regulated Genes by Chromatin Immunoprecipitation. MCF‑7 human breast cancer cell line treated with 300 nM camptothecin overnight were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. p53/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human/Mouse/Rat p53 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1355) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The p21 promoter was detected by standard PCR.
Immunocytochemistry
p53 in HeLa Human Cell Line. p53 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human/Mouse/Rat p53 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1355) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human p53 by Simple WesternTM. Simple Western lane view shows lysates of CEM human
T-lymphoblastoid cell line and HEK293T human embryonic kidney cell line, loaded at 0.2 mg/mL. A specific band was detected for p53 at approximately 59 kDa (as indicated) using 2.5 µg/mL of Goat Anti-Human/Mouse/Rat p53 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1355) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.
Knockout Validated
Western Blot Shows Human p53 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and p53 knockout HeLa cell line (KO). PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human/Mouse/Rat p53 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1355) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for p53 at approximately 53 kDa (as indicated) in the parental HeLacell line, but is not detectable in knockout HeLa cell line. GAPDH(Catalog # AF5718) is shown as a loading control.This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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 cellular decisions regarding proliferation, cell cycle checkpoints, and apoptosis. The importance of p53 is underscored by its mutation in over 50% of human cancers. Mice that lack one or both copies of p53 also showed an increased incidence of tumors, which makes the p53 deficient mouse a model system for studying cancer generation and progression.
  • 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.

6 Citations: Showing 1 - 6
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Species
Applications
Sample Type
  1. Dysfunction of the MDM2/p53 axis is linked to premature aging
    Authors: D Lessel, D Wu, C Trujillo, T Ramezani, I Lessel, MK Alwasiyah, B Saha, FM Hisama, K Rading, I Goebel, P Schütz, G Speit, J Högel, H Thiele, G Nürnberg, P Nürnberg, M Hammerschm, Y Zhu, DR Tong, C Katz, GM Martin, J Oshima, C Prives, C Kubisch
    J. Clin. Invest., 2017;0(0):.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. Growth hormone is permissive for neoplastic colon growth
    Proc Natl Acad Sci USA, 2016;0(0):.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Survivin safeguards chromosome numbers and protects from aneuploidy independently from p53.
    Authors: Wiedemuth R, Klink B, Topfer K, Schrock E, Schackert G, Tatsuka M, Temme A
    Mol Cancer, 2014;13(0):107.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. Expression of S100A6 in cardiac myocytes limits apoptosis induced by tumor necrosis factor-alpha.
    Authors: Tsoporis JN, Izhar S, Parker TG
    J. Biol. Chem., 2008;283(44):30174-83.
    Species: Rat
    Sample Type: Cell Lysates
    Application: WB
  5. Protection of human keratinocytes from UVB-induced inflammation using root extract of Lithospermum erythrorhizon.
    Authors: Ishida T, Sakaguchi I,
    Biol. Pharm. Bull., 2007;30(5):928-34.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  6. TIMP-1 gene deficiency increases tumour cell sensitivity to chemotherapy-induced apoptosis.
    Authors: Davidsen ML, Wurtz SØ, Romer MU, Sorensen NM, Johansen SK, Christensen IJ, Larsen JK, Offenberg H, Brunner N, Lademann U
    Br. J. Cancer, 2006;95(8):1114-20.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
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