Overview

  • Product nameAnti-MDM2 antibody
    See all MDM2 primary antibodies
  • Description
    Rabbit polyclonal to MDM2
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Dog, Human, African Green Monkey, Syrian Hamster
  • Immunogen

    Synthetic peptide within Human MDM2 aa 393-424 (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    Sequence:

    DYSQPSTSSSIIYSSQEDVKEFEREETQDKEE


    Database link: Q00987

  • Positive control
    • WB: Mouse lung tissue lysate and HepG2 cell lysate. ICC/IF: HeLa cells. IHC-P: Human prostate carcinoma tissue.
  • General notes

    MDM2 human protein exists in 11 isoforms UniProtKB - Q00987, the immunogen of ab38178 corresponds to atleast 10 isoforms i.e. 1,2,3,4,5,6 8, 9, 10 and 11; therefore the western blot results might show bands corresponding to these isoforms.

Properties

Applications

Our Abpromise guarantee covers the use of ab38618 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
ICC/IF 1/200.
WB 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
IHC-P 1/10 - 1/50.

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 antibody images

  • ICC/IF image of ab38618> stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 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 (ab38618, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L)(ab150077) 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.

  • Immunofluorescence analysis of MDCK cells, staining MDM2 with ab38618.

    Cells were fixed with formaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in 1% BSA in PBS) for 1 hour at 25°C. An Alexa Fluor® 488-conjugated goat anti-rabbit polyclonal IgG (1/500) (ab150077) was used as the secondary antibody.

    See Abreview

  • Anti-MDM2 antibody (ab38618) at 1/500 dilution + Mouse 3T3 Lysate at 75 µg

    Secondary
    HRP conjugated Goat anti-rabbit at 1/4000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 55 kDa


    Exposure time : 15 minutes

    This image is courtesy of an anonymous Abreview

    Blocked using 5% Milk for 30 minutes at Room temperature. Antibody Dilutent was5% milk in TBS-Tween-20 (0.05%). Primary incubation 3 hours at room temperature.

    See Abreview

  • Anti-MDM2 antibody (ab38618) + HepG2 cell lysate at 35 µg

    Predicted band size : 55 kDa
  • Anti-MDM2 antibody (ab38618) at 1/1000 dilution + Mouse lung tissue lysate at 35 µg/ml

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/5000 dilution
    Developed using the ECL technique

    Predicted band size : 55 kDa
    Observed band size : 55 kDa

    Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling MDM2 with ab38618. A peroxidase-conjugated anti-rabbit IgG was used as the secondary antibody, followed by DAB staining.

  • ab38618 staining human cancer tissue by immunohistochemistry, formalin-fixed, paraffin-embedded tissue. The hepatocarcinoma tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining.

References for Anti-MDM2 antibody (ab38618)

This product has been referenced in:
  • Rivera-Calderón LG  et al. Alterations in PTEN, MDM2, TP53 and AR protein and gene expression are associated with canine prostate carcinogenesis. Res Vet Sci 106:56-61 (2016). Read more (PubMed: 27234536) »
  • Wang J  et al. Mechanism of QSYQ on anti-apoptosis mediated by different subtypes of cyclooxygenase in AMI induced heart failure rats. BMC Complement Altern Med 15:352 (2015). WB ; Rat . Read more (PubMed: 26445960) »

See all 3 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunoprecipitation
Sample Mouse Cell lysate - whole cell (colon cancer cell line)
Total protein in input 40 µg
Immuno-precipitation step Protein A
Specification colon cancer cell line
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Submitted Oct 31 2016

Application Western blot
Sample Dog Cell lysate - whole cell (CPEK)
Gel Running Conditions Reduced Denaturing (4-20% polyacrylamide)
Loading amount 20 µg
Specification CPEK
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5%
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Submitted Apr 16 2015

Application Western blot
Loading amount 50000 cells
Gel Running Conditions Reduced Denaturing (10% gel)
Sample Human Cell lysate - whole cell (Normal prostate epithelial cells, RWPE-1)
Specification Normal prostate epithelial cells, RWPE-1
Blocking step Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 27°C
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Submitted Apr 10 2015

Application Western blot
Loading amount 60 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Mouse Tissue lysate - whole (Liver)
Specification Liver
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Oct 02 2013

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Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (NRK)
Specification NRK
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Mar 20 2013

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Mar 20 2013

Application Immunocytochemistry/ Immunofluorescence
Sample Monkey Cell (COS)
Specification COS
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Mar 20 2013

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Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (NIH3T3)
Specification NIH3T3
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Mar 20 2013

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HEK293T)
Specification HEK293T
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% triton in PBS (20 min)
Blocking step BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Nov 06 2012

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Application Immunocytochemistry/ Immunofluorescence
Sample Dog Cell (MDCK cells)
Specification MDCK cells
Fixative Formaldehyde
Permeabilization Yes - Triton (0.1%) in PBS, 3min
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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Submitted Nov 02 2012

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