Recombinant
RabMAb

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

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Overview

  • 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.

    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
    For more information see here.

    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

Applications

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.

    • Target

    • 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.
    • 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

      Images

      • Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)
        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)
        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.

      • Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880)
        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 - 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)
        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.

      References

      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) »
      • 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). Read more (PubMed: 31289362) »
      See all 6 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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