Product nameAnti-MDM2 (phospho S166) antibody [EPR1450(2)]
See all MDM2 primary antibodies
DescriptionRabbit monoclonal [EPR1450(2)] to MDM2 (phospho S166)
Tested applicationsSuitable for: WB, IHC-P, Dot blotmore details
Unsuitable for: Flow Cyt,ICC or IP
Species reactivityReacts with: Rat, Human
Does not react with: Mouse
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
- MCF-7 cell lysates treated with IGF-1, Human gastric carcinoma tissue and Human placenta tissue
Storage instructionsShipped 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 bufferPreservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
PurityTissue culture supernatant
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.
|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.|
FunctionE3 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 specificityUbiquitous. 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 diseaseNote=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 similaritiesBelongs to the MDM2/MDM4 family.
Contains 1 RanBP2-type zinc finger.
Contains 1 RING-type zinc finger.
Contains 1 SWIB domain.
DomainRegion 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.
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 localizationNucleus > 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
- ACTFS antibody
- Double minute 2 protein antibody
- E3 ubiquitin-protein ligase Mdm2 antibody
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.
All lanes : HRP labeled goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 55 kDa
Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labeling MDM2 (phospho S166) using ab170880 at a 1/50 dilution.
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.
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 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.
Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling MDM2 (phospho S166) using ab170880 at a 1/50 dilution.
This product has been referenced in:
- 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). Read more (PubMed: 30132214) »
- 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) »