Recombinant Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)
![Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)](https://www.abcam.com/ps/products/170/ab170880/Images/ab170880-171409-anti-mdm2-phospho-s166-antibody-epr14502-western-blot.jpg "Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1450(2)] to MDM2 (phospho S166)
- Suitable for: WB, IHC-P, Dot blot
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-MDM2 (phospho S166) antibody [EPR1450(2)]
See all MDM2 primary antibodies -
Description
Rabbit monoclonal [EPR1450(2)] to MDM2 (phospho S166) -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Dot blotmore details
Unsuitable for: Flow Cyt,ICC or IP -
Species reactivity
Reacts with: Human
Predicted to work with: Rat
Does not react with:
Mouse -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- MCF-7 cell lysates treated with IGF-1, Human gastric carcinoma tissue and Human placenta tissue
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR1450(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab170880 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/50000 - 1/200000. Predicted molecular weight: 55 kDa.
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| IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| Dot blot |
1/1000.
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| Notes |
|---|
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WB
1/50000 - 1/200000. Predicted molecular weight: 55 kDa. |
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IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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Dot blot
1/1000. |
Application notes
Is unsuitable for Flow Cyt,ICC or IP.
Target
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Function
E3 ubiquitin-protein ligase that mediates ubiquitination of p53/TP53, leading to its degradation by the proteasome. Inhibits p53/TP53- and p73/TP73-mediated cell cycle arrest and apoptosis by binding its transcriptional activation domain. Also acts as an 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 component of the TRIM28/KAP1-ERBB4-MDM2 complex which links growth factor and DNA damage response pathways. -
Tissue specificity
Ubiquitous. Isoform Mdm2-A, isoform Mdm2-B, isoform Mdm2-C, isoform Mdm2-D, isoform Mdm2-E, isoform Mdm2-F and isoform Mdm2-G are observed in a range of cancers but absent in normal tissues. -
Involvement in disease
Note=Seems to be amplified in certain tumors (including soft tissue sarcomas, osteosarcomas and gliomas). A higher frequency of splice variants lacking p53 binding domain sequences was found in late-stage and high-grade ovarian and bladder carcinomas. Four of the splice variants show loss of p53 binding. -
Sequence similarities
Belongs to the MDM2/MDM4 family.
Contains 1 RanBP2-type zinc finger.
Contains 1 RING-type zinc finger.
Contains 1 SWIB domain. -
Domain
Region I is sufficient for binding p53 and inhibiting its G1 arrest and apoptosis functions. It also binds p73 and E2F1. Region II contains most of a central acidic region required for interaction with ribosomal protein L5 and a putative C4-type zinc finger. The RING finger domain which coordinates two molecules of zinc interacts specifically with RNA whether or not zinc is present and mediates the heterooligomerization with MDM4. It is also essential for its ubiquitin ligase E3 activity toward p53 and itself. -
Post-translational
modificationsPhosphorylated in response to ionizing radiation in an ATM-dependent manner.
Auto-ubiquitinated; which leads to proteasomal degradation. Deubiquitinated by USP2 leads to its accumulation and increases deubiquitinilation and degradation of p53/TP53. Deubiquitinated by USP7; leading to stabilize it. -
Cellular localization
Nucleus > nucleoplasm. Cytoplasm. Nucleus > nucleolus. Expressed predominantly in the nucleoplasm. Interaction with ARF(P14) results in the localization of both proteins to the nucleolus. The nucleolar localization signals in both ARF(P14) and MDM2 may be necessary to allow efficient nucleolar localization of both proteins. Colocalizes with RASSF1 isoform A in the nucleus. - Information by UniProt
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Database links
- Entrez Gene: 4193 Human
- Entrez Gene: 314856 Rat
- Omim: 164785 Human
- SwissProt: Q00987 Human
- Unigene: 484551 Human
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Alternative names
- ACTFS antibody
- Double minute 2 protein antibody
- E3 ubiquitin-protein ligase Mdm2 antibody
see all
Images
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Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)All lanes : Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880) at 1/50000 dilution
Lane 1 : MCF7 cell lysates
Lane 2 : MCF7 cell lysates treated with IGF-1
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labeled goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 55 kDa -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)](https://www.abcam.com/ps/products/170/ab170880/Images/ab170880-171411-anti-mdm2-phospho-s166-antibody-epr14502-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labeling MDM2 (phospho S166) using ab170880 at a 1/50 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)](https://www.abcam.com/ps/products/170/ab170880/Images/ab170880-272277-anti-mdm2-phospho-s166-antibody-epr14502-western-blot.jpg)
Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)All lanes : Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880) at 1/100000 dilution
Lane 1 : A549 (Human lung carcinoma epithelial cell) whole cell lysates.
Lane 2 : A549 (Human lung carcinoma epithelial cell) treated with insulin at 1ug/ml for 150 minutes. Whole cell lysates.
Lane 3 : A549 (Human lung carcinoma epithelial cell) treated with insulin at 1ug/ml for 150 minutes. Whole cell lysates. Then the membrane was incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 55 kDa
Observed band size: 90-140 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsDiluting and blocking buffer: 2% BSA/TBST
The molecular weight is the same with the one from this paper PMID: 25392082 -
![Dot Blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)](https://www.abcam.com/ps/products/170/ab170880/Images/ab170880-270040-anti-mdm2-phospho-s166-antibody-epr14502-dot-blot.jpg)
Dot Blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)Dot blot analysis of MDM2 (phospho S166) phospho peptide (Lane 1), MDM2 non-phospho peptide (Lane 2), labeling MDM2 (phospho S166) with ab170880 at a dilution of 1/1000. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/100000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)](https://www.abcam.com/ps/products/170/ab170880/Images/ab170880-171410-anti-mdm2-phospho-s166-antibody-epr14502-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling MDM2 (phospho S166) using ab170880 at a 1/50 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)](https://www.abcam.com/ps/products/170/ab170880/Images/ab170880-6-benefits-of-recombinant-antibodies.png)
Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)
Protocols
Datasheets and documents
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Datasheet download
References (10)
ab170880 has been referenced in 10 publications.
- Machado RAC et al. L-plastin Ser5 phosphorylation is modulated by the PI3K/SGK pathway and promotes breast cancer cell invasiveness. Cell Commun Signal 19:22 (2021). PubMed: 33618712
- Zhu X et al. Overexpression of DJ-1 enhances colorectal cancer cell proliferation through the cyclin-D1/MDM2-p53 signaling pathway. Biosci Trends 14:83-95 (2020). PubMed: 32132307
- Song J et al. Downregulation of AKT and MDM2, Melatonin Induces Apoptosis in AGS and MGC803 Cells. Anat Rec (Hoboken) 302:1544-1551 (2019). PubMed: 30809951
- Zou Y et al. Chlamydia trachomatis plasmid-encoded protein Pgp3 inhibits apoptosis via the PI3K-AKT-mediated MDM2-p53 axis. Mol Cell Biochem 452:167-176 (2019). PubMed: 30132214
- Sun B et al. Stepwise detection and evaluation reveal miR-10b and miR-222 as a remarkable prognostic pair for glioblastoma. Oncogene 38:6142-6157 (2019). PubMed: 31289362
- Song J et al. Melatonin induces the apoptosis and inhibits the proliferation of human gastric cancer cells via blockade of the AKT/MDM2 pathway. Oncol Rep 39:1975-1983 (2018). PubMed: 29484412
- Kang C et al. Baicalin alleviates 6-hydroxydopamine-induced neurotoxicity in PC12 cells by down-regulation of microRNA-192-5p. Brain Res N/A:N/A (2018). PubMed: 30552896
- Cheng ZX et al. MicroRNA-132 induces temozolomide resistance and promotes the formation of cancer stem cell phenotypes by targeting tumor suppressor candidate 3 in glioblastoma. Int J Mol Med 40:1307-1314 (2017). PubMed: 28901390
- Zhang X et al. PI3K/Akt/p53 pathway inhibits reovirus infection. Infect Genet Evol 34:415-22 (2015). PubMed: 26066464
- Blanchard Z et al. BRCA1-IRIS inactivation overcomes paclitaxel resistance in triple negative breast cancers. Breast Cancer Res 17:5 (2015). PubMed: 25583261
