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Rabbit Recombinant Monoclonal MDM2 antibody. Suitable for IP, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 10 publications.


Images

Western blot - Anti-MDM2 antibody [EPR22256-98] (AB259265), expandable thumbnail
  • Immunoprecipitation - Anti-MDM2 antibody [EPR22256-98] (AB259265), expandable thumbnail
  • Western blot - Anti-MDM2 antibody [EPR22256-98] (AB259265), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-MDM2 antibody [EPR22256-98] (AB259265), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPFlow CytWBIHC-PICC/IF
Human
Tested
Not recommended
Tested
Not recommended
Tested
Mouse
Expected
Not recommended
Tested
Not recommended
Expected
Rat
Expected
Not recommended
Tested
Not recommended
Expected

Tested
Tested

Species
Human
Dilution info
1/30
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Rat
Dilution info
1/1000
Notes

-

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/100
Notes

-

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Associated Products

Select an associated product type

7 products for Alternative Product

Target data

Function

E3 ubiquitin-protein ligase that mediates ubiquitination of p53/TP53, leading to its degradation by the proteasome (PubMed:29681526). Inhibits p53/TP53- and p73/TP73-mediated cell cycle arrest and apoptosis by binding its transcriptional activation domain. Also acts as a ubiquitin ligase E3 toward itself and ARRB1. Permits the nuclear export of p53/TP53. Promotes proteasome-dependent ubiquitin-independent degradation of retinoblastoma RB1 protein. Inhibits DAXX-mediated apoptosis by inducing its ubiquitination and degradation. Component of the TRIM28/KAP1-MDM2-p53/TP53 complex involved in stabilizing p53/TP53. Also a component of the TRIM28/KAP1-ERBB4-MDM2 complex which links growth factor and DNA damage response pathways. Mediates ubiquitination and subsequent proteasome degradation of DYRK2 in nucleus. Ubiquitinates IGF1R and SNAI1 and promotes them to proteasomal degradation (PubMed:12821780, PubMed:15053880, PubMed:15195100, PubMed:15632057, PubMed:16337594, PubMed:17290220, PubMed:19098711, PubMed:19219073, PubMed:19837670, PubMed:19965871, PubMed:20173098, PubMed:20385133, PubMed:20858735, PubMed:22128911). Ubiquitinates DCX, leading to DCX degradation and reduction of the dendritic spine density of olfactory bulb granule cells (By similarity). Ubiquitinates DLG4, leading to proteasomal degradation of DLG4 which is required for AMPA receptor endocytosis (By similarity). Negatively regulates NDUFS1, leading to decreased mitochondrial respiration, marked oxidative stress, and commitment to the mitochondrial pathway of apoptosis (PubMed:30879903). Binds NDUFS1 leading to its cytosolic retention rather than mitochondrial localization resulting in decreased supercomplex assembly (interactions between complex I and complex III), decreased complex I activity, ROS production, and apoptosis (PubMed:30879903).

Alternative names

E3 ubiquitin-protein ligase Mdm2, Double minute 2 protein, Oncoprotein Mdm2, RING-type E3 ubiquitin transferase Mdm2, p53-binding protein Mdm2, Hdm2, MDM2

Recommended products

Rabbit Recombinant Monoclonal MDM2 antibody. Suitable for IP, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 10 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR22256-98
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

MDM2 also known as murine double minute 2 is an important regulator of the p53 tumor suppressor protein. It functions as an E3 ubiquitin ligase targeting p53 for proteasomal degradation. The MDM2 protein has a molecular weight of approximately 55 kDa. It is expressed in various human tissues with higher levels in rapidly dividing cells. MDM2 interacts through its N-terminal domain with the transactivation domain of p53 inhibiting its transcriptional activity.

Biological function summary

The MDM2 protein plays a central role in cell cycle control and apoptosis regulation. It forms a complex with p53 which modulates p53's stability and activity. By binding to p53 MDM2 prevents p53 from inducing cell cycle arrest or apoptosis in response to DNA damage or oncogenic signals. Its interaction allows cells to proliferate even in the presence of potential growth-arrest signals maintaining homeostasis under normal physiological conditions.

Pathways

MDM2 and p53 form a critical axis within the DNA damage response and tumorigenesis pathways. The p53-MDM2 feedback loop is a well-studied mechanism that balances cell survival and death. When DNA damage occurs p53 gets activated and in turn upregulates MDM2 expression creating a feedback loop. Other proteins like ARF also interact with MDM2 inhibiting its activity towards p53 and thereby enhancing p53 function during cellular stress.

Associated diseases and disorders

Mutations or overexpression of the MDM2 protein are often associated with various cancers including sarcoma and breast cancer. In these cancers MDM2 amplification leads to enhanced degradation of p53 resulting in unchecked cell proliferation. MDM2's involvement in these pathologies highlights its potential as a therapeutic target with inhibitors like Nutlin-3 being developed to disrupt the MDM2-p53 interaction. The relationship between MDM2 and other proteins such as HDM201 in the context of p53 alterations further highlights its importance in cancer biology.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Western blot - Anti-MDM2 antibody [EPR22256-98] (ab259265), expandable thumbnail

    Western blot - Anti-MDM2 antibody [EPR22256-98] (ab259265)

    Blocking and Dilution Buffer and concentration: 5% NFDM/TBST.

    MDM2 can be cleaved into a 60-kDa fragment after Nutlin 3a treatment as Nutlin 3a disrupts p53-MDM2 interaction and induces p53- dependent apoptosis and autophagy.

    The molecular weight observed is consistent with what has been described in the literature (PMID:19638413, 10329737).

    Exposure time: 15 seconds.

    All lanes: Western blot - Anti-MDM2 antibody [EPR22256-98] (ab259265) at 1/1000 dilution

    Lane 1: Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

    Lane 2: RAW 264.7 treated with 10uM Nutlin-3a for 24 hours, whole cell lysate at 20 µg

    Lane 3: Untreated 2.4G2 (rat B cell lymphoma B lymphocyte), whole cell lysate at 20 µg

    Lane 4: 2.4G2 treated with 10uM Nutlin-3a for 24 hours, whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 55 kDa

    Observed band size: 60 kDa, 90 kDa

  • Immunoprecipitation - Anti-MDM2 antibody [EPR22256-98] (ab259265), expandable thumbnail

    Immunoprecipitation - Anti-MDM2 antibody [EPR22256-98] (ab259265)

    MDM2 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) treated with 10uM Nutlin-3a for 24 hours whole cell lysate 10ug with ab259265 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259265 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) treated with 10uM Nutlin-3a for 24 hours whole cell lysate 10ug

    Lane 2: ab259265 IP in HepG2 treated with 10uM Nutlin-3a for 24 hours whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab259265 in HepG2 treated with 10uM Nutlin-3a for 24 hours whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

    MDM2 can be cleaved into a 60-kDa fragment after Nutlin 3a treatment as Nutlin 3a disrupts p53–MDM2 interaction and induces p53- dependent apoptosis and autophagy.

    The molecular weight observed is consistent with what has been described in the literature (PMID:19638413, 10329737).

    All lanes: Immunoprecipitation - Anti-MDM2 antibody [EPR22256-98] (ab259265)

    Predicted band size: 55 kDa

    Observed band size: 60 kDa, 90 kDa

  • Western blot - Anti-MDM2 antibody [EPR22256-98] (ab259265), expandable thumbnail

    Western blot - Anti-MDM2 antibody [EPR22256-98] (ab259265)

    Blocking and Dilution Buffer and concentration: 5% NFDM/TBST.

    MDM2 can be cleaved into a 60-kDa fragment after Nutlin 3a treatment as Nutlin 3a disrupts p53-MDM2 interaction and induces p53- dependent apoptosis and autophagy.

    The molecular weight observed is consistent with what has been described in the literature (PMID:19638413, 10329737).

    Exposure time: 10 seconds.

    All lanes: Western blot - Anti-MDM2 antibody [EPR22256-98] (ab259265) at 1/1000 dilution

    Lane 1: Untreated HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 10 µg

    Lane 2: HepG2 treated with 10uM Nutlin-3a for 24 hours, whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 55 kDa

    Observed band size: 60 kDa, 90 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-MDM2 antibody [EPR22256-98] (ab259265), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-MDM2 antibody [EPR22256-98] (ab259265)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling MDM2 with ab259265 at 1/100 (5 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear staining in HepG2 cells treated with Nutlin-3a (10 uM) for 24 hours is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab259265 anti- MDM2 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 ug/ml) dilution.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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