MDM2/HDM2 Antibody [Unconjugated]

MDM2/HDM2 was detected in immersion fixed paraffin-embedded sections of human kidney using Rabbit Anti-Human/Mouse/Rat MDM2/HDM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1244) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei for epithelial cells in convoluted tubules. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

MDM2 was detected in perfusion fixed frozen sections of mouse kidney using Human/Mouse/Rat MDM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1244) at 5 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 Frozen Tissue Sections.

Western blot shows lysates of U2OS human osteosarcoma cell line, MCF-7 human breast cancer cell line, and CEM human T-lymphoblastoid cell line untreated (-) or treated (+) with 5 µM LLnV for 1 hour. PVDF membrane was probed with 1 µg/mL of Human/Mouse/Rat MDM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1244), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for MDM2 at approximately 95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

• Reactivity: Hu, Mu, Rt
• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

MDM2/HDM2 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human MDM2
  Asn3-Pro491
  Accession # Q00987
• Specificity: Detects human, mouse, and rat MDM2.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: MDM2
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 3-15 ug/mL
  • Western Blot 1 ug/mL

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 MDM2/HDM2 Antibody [Unconjugated]

• ACTFS
• HDM2
• HDMX
• MDM2 oncogene, E3 ubiquitin protein ligase
• MDM2

Background

MDM2 is a key regulator of p53 tumor suppressor protein activity and stability. MDM2 binds to and inhibits the transactivation domain of p53. In addition, MDM2 controls p53 stability by functioning as its E3 ligase in ubiquitination and by shuttling p53 from the nucleus to the cytoplasm for subsequent degradation. The importance of the p53/MDM2 relationship is underscored by the existence of an autoregulatory feedback loop whereby activated p53 transcriptionally up‑regulates the expression of its own inhibitor, MDM2.

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 MDM2/HDM2 Antibody (AF1244)(22)

Publications using AF1244

• Nakagawa-Saito, Y;Mitobe, Y;Togashi, K;Suzuki, S;Sugai, A;Takenouchi, S;Nakamura, K;Sonoda, Y;Kitanaka, C;Okada, M; The MDM2-p53 Axis Represents a Therapeutic Vulnerability Unique to Glioma Stem Cells International journal of molecular sciences 2024-04-02 [PMID: 38612758] (Western Blot, Human)
• Ellison, V;Polotskaia, A;Xiao, G;Leybengrub, P;Qiu, W;Lee, R;Hendrickson, R;Hu, W;Bargonetti, J; A CANCER PERSISTENT DNA REPAIR CIRCUIT DRIVEN BY MDM2, MDM4 (MDMX), AND MUTANT P53 FOR RECRUITMENT OF MDC1 AND 53BP1 TO CHROMATIN bioRxiv : the preprint server for biology 2024-01-23 [PMID: 38328189] (Western Blot, Human)
• Meah, A;Vedarethinam, V;Bronstein, R;Gujarati, N;Jain, T;Mallipattu, SK;Li, Y;Wang, J; Single-Cell Spatial MIST for Versatile, Scalable Detection of Protein Markers Biosensors 2023-08-27 [PMID: 37754086] (IHC, Mouse)
• Yuta Mitobe, Shuhei Suzuki, Yurika Nakagawa-Saito, Keita Togashi, Asuka Sugai, Yukihiko Sonoda et al. Antagonizing MDM2 Overexpression Induced by MDM4 Inhibitor CEP-1347 Effectively Reactivates Wild-Type p53 in Malignant Brain Tumor Cells Cancers (Basel) 2023-08-30 [PMID: 37686602]
• Mitobe, Y;Suzuki, S;Nakagawa-Saito, Y;Togashi, K;Sugai, A;Sonoda, Y;Kitanaka, C;Okada, M; The Novel MDM4 Inhibitor CEP-1347 Activates the p53 Pathway and Blocks Malignant Meningioma Growth In Vitro and In Vivo Biomedicines 2023-07-12 [PMID: 37509605] (Western Blot, Mouse)
• AC Arcones, MR Martínez-C, R Vila-Bedma, C Yáñez, I Lladó, AM Proenza, F Mayor, C Murga Cardiac GRK2 Protein Levels Show Sexual Dimorphism during Aging and Are Regulated by Ovarian Hormones Cells, 2021-03-17;10(3):. 2021-03-17 [PMID: 33803070] (Western Blot, Mouse, Rat)
• ANUSHA THADI, WILLIAM F. MORANO, MARIAN KHALILI, BLAKE D. BABCOCK, MOHAMMAD F. SHAIKH, DESHKA S. FOSTER et al. Molecular Targeting of H/MDM-2 Oncoprotein in Human Colon Cancer Cells and Stem-like Colonic Epithelial-derived Progenitor Cells Anticancer Research 2021-01-01 [PMID: 33419797]
• Gu Xiao, George K. Annor, Kimberly Fung, Outi Keinänen, Brian M. Zeglis, Jill Bargonetti Targeting Triple Negative Breast Cancer with a Nucleus-Directed p53 Tetramerization Domain Peptide Molecular Pharmaceutics 2021-01-04 [PMID: 33289569]
• A Colmenero-, MA Gómez-Muño, I Rodríguez-, A Amador-Álv, KO Henrich, D Pascual-Va, K Okonechnik, E Rivas, F Westermann, R Pardal, FM Vega Identification of VRK1 as a New Neuroblastoma Tumor Progression Marker Regulating Cell Proliferation Cancers (Basel), 2020-11-20;12(11):. 2020-11-20 [PMID: 33233777] (Western Blot, Human)
• ANUSHA THADI, LAUREN LEWIS, EVE GOLDSTEIN, ANSHU AGGARWAL, MARIAN KHALILI, LINDSAY STEELE et al. Targeting Membrane HDM-2 by PNC-27 Induces Necrosis in Leukemia Cells But Not in Normal Hematopoietic Cells Anticancer Research 2020-09-01 [PMID: 32878773]
• Roy Xiao, Yi An, Wenda Ye, Adeeb Derakhshan, Hui Cheng, Xinping Yang et al. Dual Antagonist of cIAP/XIAP ASTX660 Sensitizes HPV(−) and HPV(+) Head and Neck Cancers To TNF alpha , TRAIL, and Radiation Therapy Clinical Cancer Research 2019-11-01 [PMID: 31266830]
• Eutteum Jeong, Eutteum Brady, José A Martina, Mehdi Pirooznia, Ilker Tunc, Rosa Puertollano The transcription factors TFE3 and TFEB amplify p53 dependent transcriptional programs in response to DNA damage eLife 2018-12-06 [PMID: 30520728]
• DL Coutu, KD Kokkaliari, L Kunz, T Schroeder Multicolor quantitative confocal imaging cytometry Nat. Methods, 2017-11-13;15(1):39-46. 2017-11-13 [PMID: 29320487] (IHC, Mouse)
• R Schreckenb, P Bencsik, M Weber, Y Abdallah, C Csonka, K Gömöri, K Kiss, J Pálóczi, J Pipis, M Sárközy, P Ferdinandy, R Schulz, KD Schlüter Adverse Effects on ?-Adrenergic Receptor Coupling: Ischemic Postconditioning Failed to Preserve Long-Term Cardiac Function J Am Heart Assoc, 2017-12-22;6(12):. 2017-12-22 [PMID: 29273639] (Western Blot, Rat)
• Daniel F. Comiskey Jr, Aishwarya G. Jacob, Brianne L. Sanford, Matías Montes, Andrew K. Goodwin, Haley Steiner et al. A novel mouse model of rhabdomyosarcoma underscores the dichotomy of MDM2-ALT1 function in vivo Oncogene 2018-01-01 [PMID: 28892044]
• Phesse , T J, Myant , K B, Cole , A M, Ridgway , R A, Pearson , H, Muncan , V, van den Brink , G R, Vousden , K H, Sears , R, Vassilev , L T, Clarke , A R, Sansom , O J Endogenous c-Myc is essential for p53-induced apoptosis in response to DNA damage in vivo. Cell Death Differ, 2014-02-28;21(6):956-66. 2014-02-28 [PMID: 24583641] (Western Blot, Mouse)
• HDM2 antagonist MI-219 (spiro-oxindole), but not Nutlin-3 (cis-imidazoline), regulates p53 through enhanced HDM2 autoubiquitination and degradation in human malignant B-cell lymphomas. J Hematol Oncol, 2012-09-18;5(0):57. 2012-09-18 [PMID: 22989009] (Western Blot, Human)
• Surget S, Chiron D, Gomez-Bougie P, Descamps G, Menoret E, Bataille R, Moreau P, Le Gouill S, Amiot M, Pellat-Deceunynck C Cell death via DR5, but not DR4, is regulated by p53 in myeloma cells. Cancer Res, 2012-06-27;72(17):4562-73. 2012-06-27 [PMID: 22738917] (Western Blot, Human)
• Liying Fan, Chandraiah Lagisetti, Carol C. Edwards, Thomas R. Webb, Philip M. Potter Sudemycins, Novel Small Molecule Analogues of FR901464, Induce Alternative Gene Splicing ACS Chemical Biology 2011-06-17 [PMID: 21344922]
• Post SM, Quintas-Cardama A, Pant V, Iwakuma T, Hamir A, Jackson JG, Maccio DR, Bond GL, Johnson DG, Levine AJ, Lozano G A high-frequency regulatory polymorphism in the p53 pathway accelerates tumor development. Cancer Cell, 2010-09-14;18(3):220-30. 2010-09-14 [PMID: 20832750] (IHC-P, Mouse)
• Abhijit Ghosh, Tina Chunyuan Chen, Yvonne L. Kapila Anoikis triggers Mdm2-dependent p53 degradation Molecular and Cellular Biochemistry 2010-10-01 [PMID: 20577896]
• Rozan LM, McDonald ER, Navaraj A, Matthew EM, Dicker DT, El-Deiry WS CARPs are ubiquitin ligases that promote MDM2-independent p53 and phospho-p53ser20 degradation. J. Biol. Chem., 2006-11-22;282(5):3273-81. 2006-11-22 [PMID: 17121812] (Western Blot, Human)

Bioinformatics

• Gene Symbol: MDM2
• Uniprot:
  • Q00987()