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Purified anti-MDM2 Antibody

Pricing & Availability
Clone
W23003F (See other available formats)
Regulatory Status
RUO
Other Names
Mouse double minute 2 homolog (MDM2), E3 ubiquitin-protein ligase MDM2, RING-type E3 ubiquitin transferase MDM2, Q00987
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
W23003F_PURE_MDM2_Antibody_1_030524
IHC staining of Purified anti-MDM2 (clone W23003F) on formalin-fixed paraffin-embedded human kidney tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., 1X (Cat. No. 928502), the tissue was incubated without (panel A) and with (panel B) 5 µg/mL of Purified anti-MDM2 (clone W23003F) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 40X objective. Scale bar: 50 µm
  • W23003F_PURE_MDM2_Antibody_1_030524
    IHC staining of Purified anti-MDM2 (clone W23003F) on formalin-fixed paraffin-embedded human kidney tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., 1X (Cat. No. 928502), the tissue was incubated without (panel A) and with (panel B) 5 µg/mL of Purified anti-MDM2 (clone W23003F) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 40X objective. Scale bar: 50 µm
  • W23003F_PURE_MDM2_Antibody_2_030524
    IHC staining of Purified anti-MDM2 (clone W23003F) on formalin-fixed paraffin-embedded human breast cancer tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., 1X (Cat. No. 928502), the tissue was incubated without (panel A) and with (panel B) 5 µg/mL of Purified anti-MDM2 (clone W23003F) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 40X objective. Scale bar: 50 µm
  • W23003F_PURE_MDM2_Antibody_3_030524
    U-2 OS cells left untreated (negative control, panels A and C) and treated with the MDM2 inhibitor called Nutlin-3a (positive control, panels B and D). This inhibitor causes the loss of negative feedback, leading to increased MDM2 levels. Cells were fixed with 4% PFA Fixation Buffer (Cat. No. 420801) for 10 minutes, permeabilized with 100% ice-cold methanol for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with 5 µg/mL of Purified anti-MDM2 (clone W23003F), followed by incubation with Alexa Fluor® 594 Goat anti-rat IgG (Cat. No. 405422) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (blue). Images were merged (panels A and B). The images were captured on a Revvity Operetta CLS™ High Content Analysis System with a 63X objective. Scale bar: 50 µm
  • W23003F_PURE_MDM2_Antibody_4_030524
    Whole cell extracts (15 µg total protein) from U-2 OS cells left untreated (-) or treated (+) with the MDM2 inhibitor Nutlin-3a. This inhibitor causes the loss of negative feedback, leading to increased MDM2 levels. Lysates were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1 µg/mL of Purified anti-MDM2 (clone W23003F) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
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634851 25 µg 152€
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634852 100 µg 384€
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Description

MDM2 (E3 ubiquitin-protein ligase MDM2) serves a pivotal role in cellular regulation by acting as a negative regulator of the tumor suppressor protein p53, a key player in safeguarding genomic integrity. MDM2 regulates p53 in two ways. First, it negatively regulates p53 by directly binding to the transcriptional activation domain and preventing p53 from interacting with DNA. Second, MDM2 functions as an E3 ubiquitin ligase to target p53 for degradation via the proteasome. By negatively regulating p53, MDM2 prevents unnecessary activation of p53-mediated cellular responses such as cell cycle arrest, DNA repair, and apoptosis. MDM2 exists in a negative feedback loop with p53, as elevated levels of p53 induce transcription of MDM2 which in turn inhibits p53 function and promotes degradation. Dysregulation of MDM2 leads to disruption of the MDM2/p53 balance and can contribute to the development of various cancers, emphasizing the importance of understanding the role of MDM2 in cancer research. Furthermore, MDM2 has functions outside p53 regulation such as chromatin regulation, complicating attempts to target the protein in cancer therapy.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Recombinant fragment of the N-terminus of human MDM2
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

IHC-P - Quality tested
ICC, WB - Verified
Recommended Usage

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 5 - 10 µg/mL is suggested. For immunocytochemistry, a concentration range of 5 - 10 μg/mL is recommended. For western blotting, the suggested use of this reagent is 0.5 - 1 µg/mL. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

For immunohistochemistry (IHC), we recommend antigen retrieval with either Tris-EDTA (10 mM Tris, 1 mM EDTA, 0.05% Tween-20, pH 9.0) or Sodium Citrate H.I.E.R. (Cat. No. 928502). For human kidney tissue, we do not recommend using Tris-EDTA.

For immunocytochemistry (ICC), we recommend fixation with Fixation Buffer (Cat. No. 420801) followed by permeabilization with either 1) 0.5% Triton-X or 2) 100% ice-cold methanol. Fixation with 100% ice-cold methanol alone is not recommended.

RRID
AB_3106147 (BioLegend Cat. No. 634851)
AB_3106147 (BioLegend Cat. No. 634852)

Antigen Details

Structure
MDM2 is a 491 amino acid protein with a predicted molecular weight of 55.2 kD.
Distribution

Primarily nuclear

Function
Ubiquitin ligase, oncogene
Interaction
p53, HIPK2, p14arf, USP7
Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cancer Biomarkers, Cell Biology, Cell Cycle/DNA Replication, Cell Death, Cell Proliferation and Viability
Antigen References
  1. Wade M, et al. 2013. Nat Rev Cancer. 13:83-96.
  2. Koo N, et al. 2022. Int J Mol Sci. 23(9):5005.
  3. Haupt Y, et al. 1997. Nature. 387:296-9.
  4. Wienken M, et al. 2017. J Mol Cell Biol. 9:74-80.
Gene ID
4193 View all products for this Gene ID
UniProt
View information about MDM2 on UniProt.org
Go To Top Version: 1    Revision Date: 04.04.2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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