Anti-MDM2抗体[SMP 14] (ab3110)

概述

  • 产品名称Anti-MDM2抗体[SMP 14]
    参阅全部 MDM2 一抗
  • 描述
    小鼠单克隆抗体[SMP 14] to MDM2
  • 特异性This antibody also recognises a peptide epitope around Thr-216 of murine MDM2 (when Thr-216 is unphosphorylated) see Zhang & Prives, 2001 for further details. SMP14 also cross reacts with some cytokeratins (6, 14 & 16). This is only a problem when working with certain epithelial cells and not fibroblasts.
  • 经测试应用适用于: Flow Cyt, WB, IHC-Fr, IP, ELISA, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide corresponding to Human MDM2 aa 154-167. Cross-linked to KLH using glutaldehyde.
    Sequence:

    C-SRPSTSSRRRAISE

  • 表位Amino acids 154 - 167.
  • 阳性对照
    • Breast carcinoma. Human Pancreas cells HeLa cells

性能

应用

Our Abpromise guarantee covers the use of ab3110 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
Flow Cyt Use 1µg for 106 cells.
WB Use at an assay dependent concentration. Predicted molecular weight: 55 kDa.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC-P Use a concentration of 1 - 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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 [SMP 14] 图像

  • IHC image of MDM2 staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab3110, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab3110 staining MDM2 in human pancreas tissue (ab29816) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded tissue sections). The sections were fixed in paraformaldehyde and subjected to heat-mediated antigen retrieval in citric buffer pH 6.0, prior to blocking with 10% serum for 1 hour at 12°C. The primary antibody was diluted 1/100 and incubated with the sample for 12 hours at 4°C. An HRP-conjugated rabbit anti-mouse polyclonal was used as the secondary antibody, diluted 1/200.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab3110 (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 (ab3110, 1µg/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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-MDM2 antibody [SMP 14] (ab3110)参考文献

This product has been referenced in:
  • Zhang P  et al. p53, MDM2, eIF4E and EGFR expression in nasopharyngeal carcinoma and their correlation with clinicopathological characteristics and prognosis: A retrospective study. Oncol Lett 9:113-118 (2015). IHC ; Human . Read more (PubMed: 25435943) »
  • Elshafey R  et al. Label-free impedimetric immunosensor for ultrasensitive detection of cancer marker Murine double minute 2 in brain tissue. Biosens Bioelectron 39:220-5 (2013). Functional Studies ; Mouse . Read more (PubMed: 22898660) »

See all 7 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: EDTA pH8, 100C, 30 minutes
Sample Human Tissue sections (sarcoma)
Specification sarcoma
Permeabilization No
Fixative Formaldehyde
Username

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Verified customer

提交于 Apr 25 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (8% gel)
Sample Human Cell lysate - whole cell (human medulloblastoma cells)
Specification human medulloblastoma cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

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提交于 Apr 18 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Mouse Tissue lysate - whole (mouse brain tissue)
Total protein in input 300 µg
Specification mouse brain tissue
Immuno-precipitation step Protein A/G
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提交于 May 09 2013

Thank you for your reply.

Based on the information that you were able to supply, I would agree with you that these bands are MDM2.

If there is anything else I can help you with, please just let me know.

Thank you for your reply.

I have attached your western blot with an arrow pointing to what I think is the correct band. I picked that band as looking at the western blot data that is on the webpage for this antibody, it appears to be recogniz...

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Thank you for your reply.

As requested, I am sending ab16895 as a free of charge replacement for ab3110. Your new order number is ******.

If you have any difficulties with this new antibody or if there is anything else I can help yo...

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Thank you for your reply.

I am actually based on out of the Eugene, Or office, or else it would be great to talk to you in person. If you would like to drop in and talk to one of my colleagues based in the Kendall Sq office, I am sure they wo...

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Thank you for contacting Abcam.

I am sorry about the issues you are having with ab3110 in western blot. The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot onmouse, rat and humansamples . If we...

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Thank you and you customer for your inquiry.

1.)I amsorry for this misunderstanding. The sequence of the rat MDM2 can be found on the Swiss Prot website here:

http://www.uniprot.org/uniprot/D3ZQ36

What my colleague wanted ...

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Thank you for your inquiry.
According to the Swiss Prot home page (http://www.uniprot.org/uniprot/Q00987) for human MDM2, the protein is ca 55kDa. There are different isoforms, but all are smaller than 55kDa.
ab3110 has been used by some of o...

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