Anti-MDM2 (phospho S166) antibody (ab58532)

Overview

  • Product nameAnti-MDM2 (phospho S166) antibody
    See all MDM2 primary antibodies
  • Description
    Rabbit polyclonal to MDM2 (phospho S166)
  • Specificityab58532 detects endogenous levels of MDM2 only when phosphorylated at serine 166.
  • Tested applicationsSuitable for: ELISA, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic phosphopeptide derived from human MDM2 around the phosphorylation site of serine 166 (A-I-SP-E-T).

  • Positive control
    • WB: COS7 cell extracts.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab58532 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
ELISA 1/5000.
WB 1/500 - 1/1000. Detects a band of approximately 85 kDa (predicted molecular weight: 55 kDa).
ICC/IF Use at an assay dependent concentration.

Target

  • 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.
  • 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 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
  • Database links
  • 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

Anti-MDM2 (phospho S166) antibody images

  • All lanes : Anti-MDM2 (phospho S166) antibody (ab58532) at 1/500 dilution

    Lane 1 : COS7 cell extract
    Lane 2 : COS7 cell extract with immunising peptide at 5 µg

    Lysates/proteins at 5 µg per lane.


    Predicted band size : 55 kDa
    Observed band size : 85 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab58532 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab58532, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-MDM2 (phospho S166) antibody (ab58532)

ab58532 has not yet been referenced specifically in any publications.

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