Anti-MDM2抗体[SMP 14] (ab3110)

Reviews (7)Q&A (9)References (15)

Key features and details

  • Mouse monoclonal [SMP 14] to MDM2
  • Suitable for: WB
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

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

  • 宿主

    Mouse

  • 经测试应用

    适用于: WBmore details

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide corresponding to Human MDM2 aa 150-250. Cross-linked to KLH using glutaldehyde.

  • 表位

    Amino acids 154 - 167.

  • 阳性对照

    • HepG2 cell lysate (treated with 10uM Nutlin-3a for 24hrs)

  • 常规说明

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

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
WB Use at an assay dependent concentration. Predicted molecular weight: 55 kDa.

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

    see all

  • 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

图片

  • Western blot - Anti-MDM2 antibody [SMP 14] (ab3110)
    Western blot - Anti-MDM2 antibody [SMP 14] (ab3110)
    All lanes : Anti-MDM2 antibody [SMP 14] (ab3110) at 2 µg/ml

    Lane 1 : HepG2 cell lysate
    Lane 2 : HepG2 treated with 10uM Nutlin-3a for 24hrs cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 55 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 2: Merged signal (red and green). Green - ab3110 observed at 75 kDa. Red - loading control, ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.

    ab3110 was shown to react with MDM2 in cell lysates subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab3110 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at 2 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

文献 (15)

发表研究结果有使用 ab3110？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab3110 被引用在 15 文献中.

  • Inoue C  et al. Renal Epithelioid Angiomyolipoma Undergoing Aggressive Clinical Outcome: The MDM2 Expression in Tumor Cells of Two Cases. Tohoku J Exp Med 247:119-127 (2019). PubMed: 30799331
  • Moghadam FH  et al. Ferulic Acid exerts concentration-dependent anti-apoptotic and neuronal differentiation-inducing effects in PC12 and mouse neural stem cells. Eur J Pharmacol 841:104-112 (2018). PubMed: 30332611
  • Chen W  et al. Overexpression of GRIM-19 accelerates radiation-induced osteosarcoma cells apoptosis by p53 stabilization. Life Sci 208:232-238 (2018). PubMed: 30005830
  • Raposo TP  et al. A refined method to study gene dosage changes in vitro using CRISPR/Cas9. J Clin Pathol N/A:N/A (2017). PubMed: 28794126
  • Li P  et al. Serum anti-MDM2 and anti-c-Myc autoantibodies as biomarkers in the early detection of lung cancer. Oncoimmunology 5:e1138200 (2016). PubMed: 27467958
View all Publications for this product

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1-10 of 16 Abreviews or Q&A

Flow Cytometry abreview for Anti-MDM2 antibody [SMP 14]

 Average
Abreviews
abreview image
Application
Flow Cytometry
Sample
Human Cell (Cell lines: HL60 and NB4)
Permeabilization
Yes - 90% methanol (diluted in PBS)
Gating Strategy
Intact cells (Event Length vs 191Ir (DNA channel) --> 191Ir vs 193Ir (DNA channel). Caspase-3 cleaved positive cells were removed by gating.
Specification
Cell lines: HL60 and NB4
Preparation
Cell harvesting/tissue preparation method: Purchased suspension cell line. In culture at 37C, 5% CO2.
Sample buffer: In culture with RPMI medium supplemented with 10% FBS, 1% Penicillin-Streptomycin and 1%L-Glutamine.
Fixation
Paraformaldehyde
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Abcam user community

Verified customer

提交于 May 02 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MDM2 [SMP 14] antibody

 Good
Abreviews
abreview image
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
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Abcam user community

Verified customer

提交于 Apr 25 2014

Western blot abreview for Anti-MDM2 [SMP 14] antibody

 Excellent
Abreviews
abreview image
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
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Abcam user community

Verified customer

提交于 Apr 18 2014

Immunoprecipitation abreview for Anti-MDM2 antibody [SMP 14]

 Good
Abreviews
abreview image
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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Abcam user community

Verified customer

提交于 May 09 2013

Question

Thanks for the help. Regarding the isoforms, I checked the literatures, please see this: http://mcb.asm.org/content/early/2006/10/23/MCB.00235-06.short

Proteins encoded by the mdm2 gene, which has a pivotal role in the regulation of

22 growth and differentiation, exist principally in human and murine cells as two

23 isoforms that migrate in gels as 75 kDa and 90 kDa proteins

So I think both green bands between 50 and 100 are MDM2s.

Please help me confirm it.

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Answer

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.

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Question

Thanks for providing me the new antibody. I am quite amazed with how fast you guys could replace it and deliver it within one day.

I performed new Western with the new AB yesterday and today. Here, I am sending you the image.

I basically get three potential bands (green). Would you please help me determine which green band is actually MDM2? Because they are all not 55kD.

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Answer

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 recognizing a band at a higher molecular weight than 55kDa.

I am not familiar with this protein, but could there be multiple isoforms of the protein and this antibody is detecting one that is slightly higher.

Please let me know if there is anything else I can help you with.

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Question

Thanks for your offer.

I would like to try ab16895. Please find my shipping address in my signature.

Our previous PO number on this antibody is *******.

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Answer

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 you with, please let me know.

Read More

Question

Thanks for your prompt reply. My samples are from human melanoma cell lines.

For your questions, I loaded 150 ug per lane and the antibody dilution is 1:200.

Here, I am sending you the images of my WB of MDM2 and p53 for the same samples. The red bands are Actin as controls. The green bands are supposed to be either P53 or MDM2 respectively.

I would like to try another antibody from your company as I always like to use your antibodies.

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Answer

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 would be happy to meet you.

I agree with your assessment of theMDM2 antibody and therefore I would be very happy to offer you a replacement antibody of your choice. Some of our other MDM2 antibodies that should work in human cells in western blot are:

https://www.abcam.com/MDM2-antibody-2A10-ab16895.html
https://www.abcam.com/MDM2-antibody-ab58530.html



Please let me know which one you would like to receive and I will arrange for it to be sent to you, or have it available for you to pick up.

I hope your day is going well.

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Question

I ordered this antibody in March (Anti-MDM2 antibody [SMP 14] ab3110). It turned out no alleged band of MDM2 detected.

I am pretty good at antibody optimization, no don’t let me add more protein or use more antibody (I’ve already tried).

I just want to know, you have 4 antibodies against MDM2, which one is the best for WB detection of it?

If you could give me some clue, it will be greatly appreciated.

Read More
Answer

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 cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

I know that you tried various protein and antibody concentrations, but I was wondering if you could provide that information, along with the species of the sample that you are loading. Just so we have that information for our records.

Also, as we have a number of antibodies against MDM2, once I know which species of sample you are using, I will be able to help select the appropriate antibody for you.

I look froward to your reply.

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Question

One of my costumers wants to use anti-MDM2, SMP14 (code. ab3110) in WB assays with rat specimen.

Data Sheet says that clone SMP14 cross reacts with rat, but reading the answers for other costumers' technical doubts at ABCAM site, now I have questions:


1-) If there is no information about the human MDM2 sequence detected by ab3110 in rat, how is it possible to affirm that it crossreacts with samples of this animal?



2-) Is there any internal study, article or publication showing the use of ab3110 with success for rat samples staining?

I'm forward to hearing from you.

Thanks for your attention.

Best Regards,

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Answer

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 to say is that there is not much information to be found about the rat proteinand also isoforms are not know so far. But according to Swiss Prot site this protein is not well characterised in rat and is still in the "predicted" status. I up-dated the comment on our datasheet to avoid further confusion.

The good news is that the immunogen of ab3110 and the rat MDM2 share a 100% homology and therefore the antibody will cross-react. Since this is stated on the datasheet we will guarantee this for your customer.

2.) We currently do not know of a publication that used ab3110 in rat.

I hope this information is helpful. Please do not hesitate to contact me again with any further questions.

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