Name: Anti-MDM2 (phospho S166) antibody [EPR1450(2)]
Brand: RabMAb
SKU: ab170880

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: Rat, Human
Does not react with: Mouse
Immunogen

Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human MDM2 aa 150-250 (phospho S166) (Cysteine residue). The exact sequence is proprietary.
Database link: Q00987
Positive control
- MCF-7 cell lysates treated with IGF-1, Human gastric carcinoma tissue and Human placenta tissue
General notes

A trial size is available to purchase for this antibody.
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 rabbit monoclonal antibody.

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

Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR1450(2)
Isotype

IgG
Research areas

Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- MCF7 whole cell lysate (ab3871)

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.
IHC-P		1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Dot blot		1/1000.

Application notes

Is unsuitable for Flow Cyt,ICC or IP.

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
modifications

Phosphorylated 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.
Target information above from: UniProt accession Q00987 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 4193 Human
- Entrez Gene: 314856 Rat
- Omim: 164785 Human
- SwissProt: Q00987 Human
- Unigene: 484551 Human
- Unigene: 91829 Rat
Alternative names
- ACTFS antibody
- Double minute 2 protein antibody
- E3 ubiquitin-protein ligase Mdm2 antibody
- Hdm 2 antibody
- Hdm2 antibody
- HDMX antibody
- MDM 2 antibody
- MDM2 antibody
- MDM2 oncogene E3 ubiquitin protein ligase antibody
- Mdm2 p53 E3 ubiquitin protein ligase homolog antibody
- Mdm2 transformed 3T3 cell double minute 2 p53 binding protein (mouse) binding protein 104kDa antibody
- MDM2_HUMAN antibody
- MDM2BP antibody
- Mouse Double Minute 2 antibody
- MTBP antibody
- Murine Double Minute Chromosome 2 antibody
- Oncoprotein Mdm2 antibody
- p53 Binding Protein Mdm2 antibody
- p53-binding protein Mdm2 antibody
- Ubiquitin protein ligase E3 Mdm2 antibody
- Ubiquitin protein ligase E3 Mdm2 antibody
see all

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)

Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labeling MDM2 (phospho S166) using ab170880 at a 1/50 dilution.
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 seconds

Diluting 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)

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.
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.

Western blot protocols
Immunohistochemistry protocols

Datasheet

This product has been referenced in:

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). Read more (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). Read more (PubMed: 28901390) »

See all 4 Publications for this product

Publishing research using ab170880? Please let us know so that we can cite the reference in this datasheet.

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Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)

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