AB259275

Anti-MDM2 antibody [EPR22256-98] - BSA and Azide free

RabMAb
Recombinant
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(1 Publication)

Rabbit Recombinant Monoclonal MDM2 antibody. Carrier free. Suitable for IP, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

View 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

2 Images

Immunocytochemistry/ Immunofluorescence - Anti-MDM2 antibody [EPR22256-98] - BSA and Azide free (AB259275)

ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-MDM2 antibody [EPR22256-98] - BSA and Azide free (AB259275)

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 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. 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 ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 (2 ug/ml) dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab259265).

Unknown

Immunoprecipitation - Anti-MDM2 antibody [EPR22256-98] - BSA and Azide free (AB259275)

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) (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 (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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab259265).

All lanes:

Immunoprecipitation - Anti-MDM2 antibody [EPR22256-98] (<a href='/en-us/products/primary-antibodies/mdm2-antibody-epr22256-98-ab259265'>ab259265</a>)

Predicted band size: 55 kDa

Observed band size: 60 kDa,90 kDa

false

Unconjugated

Anti-MDM2 antibody [EPR22256-98]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR22256-98

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

WB, ICC/IF, IP

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "guaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" }, "Mouse": { "IP-species-checked": "predicted", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "guaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" }, "Rat": { "IP-species-checked": "predicted", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "guaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" } } }

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Product details

ab259275 is the carrier-free version of ab259265.

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

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

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Supplementary information

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

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.

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.

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Product protocols

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

Visit the General protocols
Visit the Troubleshooting

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Target data

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).

See full target information MDM2

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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Human molecular genetics 30:2347-2361 PubMed34270708

2021

Cardiomyocyte-produced miR-339-5p mediates pathology in Duchenne muscular dystrophy cardiomyopathy.

Applications

Unspecified application

Species

Unspecified reactive species

Melanie Gartz,Margaret Beatka,Mariah J Prom,Jennifer L Strande,Michael W Lawlor

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-MDM2 antibody [EPR22256-98] - BSA and Azide free

Immunocytochemistry/ Immunofluorescence - Anti-MDM2 antibody [EPR22256-98] - BSA and Azide free (AB259275)

Immunoprecipitation - Anti-MDM2 antibody [EPR22256-98] - BSA and Azide free (AB259275)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (1)

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