概述

  • 产品名称Anti-MDM2抗体[2A10]
    参阅全部 MDM2 一抗
  • 描述
    小鼠单克隆抗体[2A10] to MDM2
  • 特异性Recognizes the ~90 kDa (apparent MW) MDM2 protein in A549 cells.
  • 经测试应用适用于: ICC/IF, WB, IP, IHC-Fr, IHC-P, ICC, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Full length MDM2 protein (Human)

  • 表位Within amino acids 294-339.
  • 阳性对照
    • A549 cell lysates. HeLa cells

性能

应用

Our Abpromise guarantee covers the use of ab16895 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/200. Use paraformaldehyde fixed cells.
WB Use a concentration of 2 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 55 kDa).
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IHC-P Use a concentration of 2.5 µg/ml.
ICC Use a concentration of 1 µg/ml.
Flow Cyt Use 0.5µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

靶标

  • 功能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.
  • 组织特异性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.
  • 疾病相关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.
  • 序列相似性Belongs to the MDM2/MDM4 family.
    Contains 1 RanBP2-type zinc finger.
    Contains 1 RING-type zinc finger.
    Contains 1 SWIB domain.
  • 结构域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.
  • 翻译后修饰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.
  • 细胞定位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
  • 数据库链接
  • 别名
    • 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 [2A10] 图像

  • ab16895 staining MDM2 in MCF7 cells treated with dienogest (ab141282), by ICC/IF. Decrease in MDM2 expression correlates with increased concentration of dienogest, as described in literature.
    The cells were incubated at 37°C for 24 hour in media containing different concentrations of ab141282 (dienogest) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab16895 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • Anti-MDM2 antibody [2A10] (ab16895) at 1/1000 dilution + Mouse Liver lysate at 40 µg with Milk, 2 hours at 25°C at 5 %

    Secondary
    Donkey anti-mouse IgG (HRP) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 55 kDa
    Additional bands at : 55 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute

    Image is courtesy of an anonymous Abreview

    See Abreview

  • ab16895 staining MDM2 in MCF7 cells treated with progesterone (ab141252), by ICC/IF. Decrease in MDM2 expression correlates with increased concentration of progesterone, as described in literature.
    The cells were incubated at 37°C for 24 hour in media containing different concentrations of ab141252 (progesterone) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab16895 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • ab16895 staining Human normal tonsil. Staining is localised to nuclear + cytoplasmic compartments. Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.

    Sections were stained using an automated system DAKO Autostainer Plus, at RT: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 min. They were then blocked with Dako Protein block for 10 min (containing casein 0.25% in PBS) , incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 min. Colorimetric detection was completed with DAB for 5 min. Slides were counterstained with Haematoxylin and coverslipped under DePeX.

  • Anti-MDM2 antibody [2A10] (ab16895) at 2 µg/ml + A549 cell lysate

    Predicted band size : 55 kDa
    Observed band size : 90 kDa (why is the actual band size different from the predicted?)
  • Overlay histogram showing HeLa cells stained with ab16895 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum (ab7481) / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16895, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • ab16895 (1/200) staining MDM2 in assynchronous HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). Please refer to Abreview for further experimental details.

    See Abreview

Anti-MDM2 antibody [2A10] (ab16895)参考文献

This product has been referenced in:
  • Avallone G  et al. Histological Classification and Immunohistochemical Evaluation of MDM2 and CDK4 Expression in Canine Liposarcoma. Vet Pathol 53:773-80 (2016). Read more (PubMed: 26993784) »
  • Mikawa T  et al. Senescence-inducing stress promotes proteolysis of phosphoglycerate mutase via ubiquitin ligase Mdm2. J Cell Biol 204:729-45 (2014). Read more (PubMed: 24567357) »

See all 20 Publications for this product

Product Wall

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing (10% SDS PAGE)
Sample Mouse Tissue lysate - whole (Liver)
Specification Liver
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jun 02 2014

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Immuno-precipitation step Protein A/G
Sample Mouse Tissue lysate - whole (2-wk-old mouse brain)
Specification 2-wk-old mouse brain
Total protein in input 200 µg
Username

Abcam user community

Verified customer

提交于 Jun 12 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Loading amount 20 µg
Specification HEK293
Treatment lane 2, CHX 4 hrs
Gel Running Conditions Reduced Denaturing (8%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 01 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts)
Loading amount 20 µg
Specification Mouse embryonic fibroblasts
Gel Running Conditions Reduced Denaturing (4-20%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jan 23 2013

Merci de nous avoir contactés.

Je ne pense pas que la bande à 45 kDa puisse correspondre à la forme non-modifiée de la Mdm2 humaine, isoforme de poids moléculaire 55kDa, http://www.uniprot.org/uniprot/Q00987....

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Thank you for your phone call.

The increased molecular weight is likely due to ubiquitinization. According to the UniProt database (Q00987), MDM2 is auto-ubiquitinated; which leads to proteasomal degradation. It is also ubiquitinated by TRIM1...

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Thank you for contacting us.

ab95934 is recombinant full length Human MDM2 Isoform 11. The difference between isoform 1 and isoform 11 is that isoform 11 has 6 more amino acid (VRSRQM) before a.a 2 -491 of isoform 1. As the epitope recognize...

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Thank you for contacting us. Here is the link to the Uniprot entry for the human Mdm2 protein: http://www.uniprot.org/uniprot/Q00987 I hope this information is helpful to you. Please do not hesitate to contact me if you need any more advice or informat...

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Thank you for contacting us. It was nice talking to you. As discussed please find attached all the information of anti MSDM2 antibodies in Excel format. I hope this information is helpful to you. Please do not hesitate to contact us if yo...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100
Username

Dr. Kirk McManus

Verified customer

提交于 Aug 03 2011

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