Overview

  • Product name

    Anti-MDM2 antibody - ChIP Grade
    See all MDM2 primary antibodies
  • Description

    Rabbit polyclonal to MDM2 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, ChIPmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Cow, Cat, Dog, Pig, Rhesus monkey
  • Immunogen

    Recombinant fragment within Human MDM2 (internal sequence). The exact sequence is proprietary.
    Database link: Q00987

  • Positive control

    • WB: Neuro-2a, GL261, C8D30, NIH/3T3, RAW 264.7, BCL-1, C2C12, Jurkat, Raji and NCIH-929 whole cell extracts; MDM2 transfected HEK-293T whole cell extract; HepG2, starved for 16 hours, whole cell extract. IP: Jurkat whole cell extract. IHC-P: Human lung and lung cancer tissues. ChIP: PC-3 chromatin extract.

Properties

Applications

Our Abpromise guarantee covers the use of ab226939 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 1/500 - 1/3000. Predicted molecular weight: 55 kDa.
IP 1/100 - 1/500.
IHC-P 1/100 - 1/1000.
ChIP Use at an assay dependent concentration.

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

  • All lanes : Anti-MDM2 antibody - ChIP Grade (ab226939) at 1/2000 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell extract
    Lane 2 : MDM2 transfected HEK-293T whole cell extract

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Predicted band size: 55 kDa



    7.5% SDS-PAGE gel.

  • MDM2 was immunoprecipitated from Jurkat (human T cell leukemia cell line from peripheral blood) whole cell extract with 4 µg ab226939. Western blot was performed from the immunoprecipitate using ab226939  at 1/500 dilution.

    Lane 1: Jurkat whole cell extract 30 μg (Input).

    Lane 2: Control IP in Jurkat whole cell extract with 4 μg of preimmune Rabbit IgG.

    Lane 3: ab226939 IP in Jurkat whole cell extract.

    7.5 % SDS-PAGE gel.

  • All lanes : Anti-MDM2 antibody - ChIP Grade (ab226939) at 1/1000 dilution

    Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell extract
    Lane 2 : Raji (human Burkitt's lymphoma cell line) whole cell extract
    Lane 3 : NHI-H929 whole cell extract

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 55 kDa



    7.5% SDS-PAGE gel.

  • All lanes : Anti-MDM2 antibody - ChIP Grade (ab226939) at 1/1000 dilution

    Lane 1 : Neuro-2a (mouse neuroblastoma cell line) whole cell extract
    Lane 2 : GL261 whole cell extract
    Lane 3 : C8D30 whole cell extract
    Lane 4 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell extract
    Lane 5 : BCL-1 whole cell extract
    Lane 6 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell extract
    Lane 7 : C2C12 (mouse myoblast cell line) whole cell extract

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Predicted band size: 55 kDa



    7.5% SDS-PAGE gel.

  • Paraffin-embedded human lung cancer tissue stained for MDM2 with ab226939 at 1/500 dilution in immunohistochemical analysis.

  • Paraffin-embedded human lung tissue stained for MDM2 with ab226939 at 1/500 dilution in immunohistochemical analysis.

  • All lanes : Anti-MDM2 antibody - ChIP Grade (ab226939) at 1/3000 dilution

    Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line), starved for 16 hours, whole cell extract
    Lane 2 : HepG2, starved for 16 hours and treated with 1 uM staurosporine for 24 hours, whole cell extract

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG

    Predicted band size: 55 kDa



    7.5% SDS-PAGE gel.

  • Cross-linked ChIP was performed with PC-3 (human prostate adenocarcinoma cell line) chromatin extract and 5 μg of either control rabbit IgG or ab226939. The precipitated DNA was detected by PCR with primer set targeting to PSA promoter.

References

This product has been referenced in:

See 1 Publication for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (A549, PC9 (lung cancer cell line))
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
12 µg
Specification
A549, PC9 (lung cancer cell line)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 01 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (MDA-MB-231 cell)
Permeabilization
Yes - Tween-20
Specification
MDA-MB-231 cell
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Oct 29 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (glioblastoma tissue)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 6.0 Citric Acid
Permeabilization
Yes - Tween-20
Specification
glioblastoma tissue
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 29 2018

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