p53 Antibody [Unconjugated]

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.

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.

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.

Simple Western lane view shows lysates of CEM humanT-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 the12-230 kDa separation system.

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 HeLa cell 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.

Overexpression of Survivin in glioma cells affects cell cycle and proliferation in a p53 dependent manner. a Western blot analysis showing cell lysates from U87-MG and U87-MGshp53 after transduction with Survivin and control vectors. The membranes were blotted with anti-p53 (53 kDa), anti-p53s15 (53 kDa), anti-p21waf/cip (21 kDa), anti-gamma H2AX (16 kDa), anti-cyclin D1 (37 kDa) and E (42 kDa singlets +50 kDa doublets) antibodies and the signals comparing control and Survivin cells were measured. The relative band density (fold increase) obtained from the densitometric analysis and normalized to the corresponding control is depicted. alpha -actin (42 kDa) was used as an internal loading control. b Representative BrdU-incorporation assays of U87-MG (left panel) and U87-MGshp53 cells (right panel) with overexpression of Survivin (bottom) compared to the empty vector-transduced controls (top). The overall amount of BrdU-positive fractions as well as the relative levels of BrdU+ and BrdU− cells in fractions with DNA contents of 2n, 4n, 8n and >8n are depicted. c Proliferation index of the U87-MG and U87-MGshp53 cells transduced with Survivin and empty vector controls. Depicted are mean values ± SD. **p 4n. Depicted are mean values ± SD. **p < 0.01. All data were collected 72 h after transduction of cells Image collected and cropped by CiteAb from the following publication (//bmccancer.biomedcentral.com/articles/10.1186/s12885-017-3932-y), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, IP, ChIP, ICC/IF, KO
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

p53 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human p53
  Asp7-Asp393
  Accession # P04637
• Specificity: Detects human, mouse, and rat p53.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: TP53
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 5 ug/5 x 10^6 cells
  • Immunocytochemistry 5-15 ug/mL
  • Immunoprecipitation 1-2 ug/500 ug cell lysate
  • Knockout Validated
  • Simple Western 2.5 ug/mL
  • Western Blot 0.25 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for p53 Antibody [Unconjugated]

• Antigen NY-CO-13
• FLJ92943
• LFS1TRP53
• p53 tumor suppressor
• p53
• P53cellular tumor antigen p53
• Phosphoprotein p53
• transformation-related protein 53
• tumor protein p53
• Tumor suppressor p53

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for p53 Antibody (AF1355)(30)

Publications using AF1355

• Oppezzo, A;Monney, L;Kilian, H;Slimani, L;Maczkowiak-Chartois, F;Rosselli, F; Fanca deficiency is associated with alterations in osteoclastogenesis that are rescued by TNF? Cell & bioscience 2023-06-24 [PMID: 37355617] (Western Blot, Mouse)
• Matsumoto C, Sekine H, Zhang N et al. Role of p53 in Cisplatin-Induced Myotube Atrophy International Journal of Molecular Sciences 2023-05-24 [PMID: 37298128]
  
  Details:
  Dilution: 10 ?g/mL
• T Nakayama, M Sunaoshi, Y Shang, M Takahashi, T Saito, BJ Blyth, Y Amasaki, K Daino, Y Shimada, A Tachibana, S Kakinuma Calorie restriction alters the mechanisms of radiation-induced mouse thymic lymphomagenesis PLoS ONE, 2023-01-20;18(1):e0280560. 2023-01-20 [PMID: 36662808] (Simple Western, Mouse)
• Maria Grazia Filippone, Stefano Freddi, Silvia Zecchini, Silvia Restelli, Ivan Nicola Colaluca, Giovanni Bertalot et al. Aberrant phosphorylation inactivates Numb in breast cancer causing expansion of the stem cell pool Journal of Cell Biology 2022-12-05 [PMID: 36200956]
• Czarnek M, Sarad K, Kara? A et al. Non-targeting control for MISSION shRNA library silences SNRPD3 leading to cell death or permanent growth arrest Molecular Therapy - Nucleic Acids 2021-12-03 [PMID: 34703654]
• EM Haberl, R Pohl, L Rein-Fisch, M Höring, S Krautbauer, G Liebisch, C Buechler Accumulation of cholesterol, triglycerides and ceramides in hepatocellular carcinomas of diethylnitrosamine injected mice Lipids in Health and Disease, 2021-10-10;20(1):135. 2021-10-10 [PMID: 34629057] (Western Blot, Mouse)
• A Rodríguez, K Zhang, A Färkkilä, J Filiatraul, C Yang, M Velázquez, E Furutani, DC Goldman, B García de, G Garza-Mayé, K McQueen, LA Sambel, B Molina, L Torres, M González, E Vadillo, R Pelayo, WH Fleming, M Grompe, A Shimamura, S Hautaniemi, J Greenberge, S Frías, K Parmar, AD D'Andrea MYC Promotes Bone Marrow Stem Cell Dysfunction in Fanconi Anemia Cell Stem Cell, 2020-09-29;0(0):. 2020-09-29 [PMID: 32997960] (Flow Cytometry, Human)
• Bailu Xie, Jan Hänsel, Vanessa Mundorf, Janina Betz, Irene Reimche, Mert Erkan et al. Pseudopterosin and O-Methyltylophorinidine Suppress Cell Growth in a 3D Spheroid Co-Culture Model of Pancreatic Ductal Adenocarcinoma Bioengineering (Basel) 2020-06-14 [PMID: 32545910]
• E Toufektcha, V Lejour, R Durand, N Giri, I Draskovic, B Bardot, P Laplante, S Jaber, BP Alter, JA Londono-Va, SA Savage, F Toledo Germline mutation of MDM4, a major p53 regulator, in a familial syndrome of defective telomere maintenance Sci Adv, 2020-04-10;6(15):eaay3511. 2020-04-10 [PMID: 32300648] (Western Blot, Mouse)
• Balázs Besztercei, Tamás Vancsik, Anett Benedek, Enikő Major, Mbuotidem J. Thomas, Csaba A. Schvarcz et al. Stress-Induced, p53-Mediated Tumor Growth Inhibition of Melanoma by Modulated Electrohyperthermia in Mouse Models without Major Immunogenic Effects International Journal of Molecular Sciences 2019-08-17 [PMID: 31426515]
• Tamas Vancsik, Gertrud Forika, Andrea Balogh, Eva Kiss, Tibor Krenacs Modulated electro‐hyperthermia induced p53 driven apoptosis and cell cycle arrest additively support doxorubicin chemotherapy of colorectal cancer in vitro Cancer Medicine 2019-08-01 [PMID: 31183995]
• Jianjun Wang, Yongliang Yao, Qinghui Zhang, Shasha Li, Lijun Tang Inflammatory responses induced by Helicobacter pylori on the carcinogenesis of gastric epithelial GES‑1 cells International Journal of Oncology 2019-04-09 [PMID: 31081048]
• ZP You, MJ Yu, YL Zhang, K Shi Progranulin protects the mouse retina under hypoxic conditions via inhibition of the Toll?like receptor?4?NADPH oxidase 4 signaling pathway Mol Med Rep, 2018-11-08;0(0):. 2018-11-08 [PMID: 30431085] (Western Blot, Mouse)
• Jin Jin, Lauren Richardson, Samantha Sheller-Miller, Nanbert Zhong, Ramkumar Menon Oxidative stress induces p38MAPK-dependent senescence in the feto-maternal interface cells Placenta 2018-07-01 [PMID: 29941169] (Western Blot, Human)
• J Wang, G He, Q Yang, L Bai, B Jian, Q Li, Z Li B-cell lymphoma 2 is associated with advanced tumor grade and clinical stage, and reduced overall survival in young Chinese patients with colorectal carcinoma Oncol Lett, 2018-04-13;15(6):9009-9016. 2018-04-13 [PMID: 29844818] (IHC-P, Western Blot, Human)
• Hongran Wang, Shuying Zhao, Michelle Barton, Todd Rosengart, Austin J. Cooney Reciprocity of Action of Increasing Oct4 and Repressing p53 in Transdifferentiation of Mouse Embryonic Fibroblasts into Cardiac Myocytes Cellular Reprogramming 2018-02-08 [PMID: 29412738] (Western Blot, Mouse)
• M Conde, S Michen, R Wiedemuth, B Klink, E Schröck, G Schackert, A Temme Chromosomal instability induced by increased BIRC5/Survivin levels affects tumorigenicity of glioma cells BMC Cancer, 2017-12-28;17(1):889. 2017-12-28 [PMID: 29282022] (Western Blot, Human)
• 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 Dysfunction of the MDM2/p53 axis is linked to premature aging J. Clin. Invest., 2017-08-28;0(0):. 2017-08-28 [PMID: 28846075] (Western Blot, Human)
• Mercy Pawar, Boris Busov, Aaruran Chandrasekhar, Jingyu Yao, David N Zacks, Cagri G Besirli FAS apoptotic inhibitory molecule 2 is a stress-induced intrinsic neuroprotective factor in the retina Cell Death & Differentiation 2017-10-01 [PMID: 28708137]
• Syeda MM, Upadhyay K, Loke J et al. Prediction of breast cancer risk based on flow-variant analysis of circulating peripheral blood B cells. Genet Med 3/16/2017 [PMID: 28301456]
• Katrien Van Roosbr Combining anti-miR-155 with chemotherapy for the treatment of lung cancers Clin. Cancer Res, 2016-11-30;0(0):. 2016-11-30 [PMID: 27903673] (Western Blot, Human)
• Growth hormone is permissive for neoplastic colon growth Proc Natl Acad Sci USA, 2016-05-25;0(0):. 2016-05-25 [PMID: 27226307] (Western Blot, Human)
• QIANG ZHOU, WENBING HU, XINXIONG FEI, XUQUN HUANG, XI CHEN, DEQING ZHAO et al. Recombinant human neuregulin-1 beta is protective against radiation-induced myocardial cell injury Molecular Medicine Reports 2016-07-01 [PMID: 27150576]
• Wiedemuth R, Klink B, Topfer K, Schrock E, Schackert G, Tatsuka M, Temme A Survivin safeguards chromosome numbers and protects from aneuploidy independently from p53. Mol Cancer, 2014-05-09;13(0):107. 2014-05-09 [PMID: 24886358] (Western Blot, Human)
• Shashi Prasad, Virginia Cody, Jennifer K. Saucier-Sawyer, Tarek R. Fadel, Richard L. Edelson, Martin A. Birchall et al. Optimization of Stability, Encapsulation, Release, and Cross-Priming of Tumor Antigen-Containing PLGA Nanoparticles Pharmaceutical Research 2012-09-01 [PMID: 22798259]
• Lucia Napione, Marina Strasly, Claudia Meda, Stefania Mitola, Maria Alvaro, Gabriella Doronzo et al. IL-12-dependent innate immunity arrests endothelial cells in G0-G1 phase by a p21(Cip1/Waf1)-mediated mechanism Angiogenesis 2012-12-01 [PMID: 22797886]
• Shashi Prasad, Virginia Cody, Jennifer K. Saucier-Sawyer, W. Mark Saltzman, Clarence T. Sasaki, Richard L. Edelson et al. Polymer nanoparticles containing tumor lysates as antigen delivery vehicles for dendritic cell-based anti-tumor immunotherapy Nanomedicine: Nanotechnology, Biology and Medicine 2011-02-01 [PMID: 20692374]
• Tsoporis JN, Izhar S, Parker TG Expression of S100A6 in cardiac myocytes limits apoptosis induced by tumor necrosis factor-alpha. J. Biol. Chem., 2008-08-27;283(44):30174-83. 2008-08-27 [PMID: 18753141] (Western Blot, Rat)
• Ishida T, Sakaguchi I Protection of human keratinocytes from UVB-induced inflammation using root extract of Lithospermum erythrorhizon. Biol. Pharm. Bull., 2007-05-01;30(5):928-34. 2007-05-01 [PMID: 17473437] (Western Blot, Human)
• Davidsen ML, Wurtz SØ, Romer MU, Sorensen NM, Johansen SK, Christensen IJ, Larsen JK, Offenberg H, Brunner N, Lademann U TIMP-1 gene deficiency increases tumour cell sensitivity to chemotherapy-induced apoptosis. Br. J. Cancer, 2006-10-23;95(8):1114-20. 2006-10-23 [PMID: 17047657] (Western Blot, Mouse)

Review for p53 Antibody (AF1355) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 01/05/2015

Summary

• Application: Western Blot
• Sample Tested: See PMID 24886358
• Species: Human

Bioinformatics

• Gene Symbol: TP53
• Uniprot:
  • P04637()