SKU PA1378-1
Size 100μg/vial
Reactivity Human
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications WB

Overview

Product Name Anti-MDM2 Antibody
SKU/Catalog Number PA1378-1
Storage & Handling At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for E3 ubiquitin-protein ligase Mdm2(MDM2) detection. Tested with WB in Human.
Cite This Product Anti-MDM2 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1378-1)
Host Rabbit
Contents/Buffer Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
Form Lyophilized
Immunogen A synthetic peptide corresponding to a sequence at the C-terminus of human MDM2(343-358aa EKAKLENSTQAEEGFD), different from the related mouse sequence by two amino acids, and from the related rat sequence by three amino acids.
Reactivity Human

Assay Details

Assay Dilutions Overview

Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Western blot, 0.1-0.5μg/ml, Human

Boster's Secondary Antibodies And IHC, WB Kits

The following reagents are used to generate the images below.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.

Images And Assay Conditions

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Figure 1. Western blot analysis of MDM2 using anti- MDM2 antibody (PA1378-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: Human Caco-2 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- MDM2 antigen affinity purified polyclonal antibody (Catalog # PA1378-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MDM2 at approximately 90KD. The expected band size for MDM2 is at 55KD.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id Q00987
Gene Name MDM2
Protein Name E3 ubiquitin-protein ligase Mdm2
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.
Alternative Names E3 ubiquitin-protein ligase Mdm2;6.3.2.-;Double minute 2 protein;Hdm2;Oncoprotein Mdm2;p53-binding protein Mdm2;MDM2;
Subcellular 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.
Molecular Weight 55233 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein 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 a 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. Mediates ubiquitination and subsequent proteasome degradation of DYRK2 in nucleus. Ubiquitinates IGF1R and SNAI1 and promotes them to proteasomal degradation. .
Research Areas Cancer, Cell Biology, Cell Cycle, Cell Cycle Inhibitors, Epigenetics And Nuclear Signaling, Oncoproteins, Oncoproteins/Suppressors, P53 Pathway, Transcription, Ubiquitin & Ubiquitin Like Modifiers

*You can search these to find other products in these research areas.
Background Mdm2 is an important negative regulator of the p53 tumor suppressor. It is the name of a gene as well as the protein encoded by that gene. Mdm2 protein functions both as an E3 ubiquity lipase that recognizes the N-terminal trans-activation domain(TAD) of the p53 tumor suppressor and an inhibitor of p53 transcriptional activation. Oliner et al.(1992) used MDM2 clones to localize the human gene to chromosome 12q13-q14 by analysis of human-hamster somatic cell hybrids.

Other Recommended Resources

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Order Product (PA1378-1)

Promotion:

Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
Option Price
30ug sample size $99
100ug $240
100ug+Free HRP Secondary BA1054 $240
100ug+Free Biotin Secondary BA1003 $240

USD $240

Ships in 7-10 business days.

Troubleshooting

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Customer Reviews

Review by
Rating:

Source: Biocompare.com

"Boster's Anti-MDM2 Antibody Worked!"

--Stacy Wilson, Biology, REVOLUTION Medicines, Inc., SRA

Applications Western Blot
Sample MDA-MB-468 and HEK293E cell lysates
Detection SuperSignal West Pico Plus with a BioRad imager

"I've been looking at downstream products of AKT and mTOR transcriptional regulation. I tried several anti-MDM2 antibodies from other companies with either ambiguous or negative results (in Western blots of human cancer cell line lysates) and Boster's is the only one that gave me strong bands at the two expected sizes. I would recommend this product. I would absolutely buy it again."

(Posted on 8/21/2017)
Write a review for PA1378-1

Publications

The Oncoprotein HBXIP Modulates the Feedback Loop of MDM2/p53 to Enhance the Growth of Breast Cancer
Liao S, Xiao S, Zhu G, Zheng D, He J, Pei Z, Li G, Zhou Y. Oncol Rep. 2014 Dec;32(6):2703-9. Doi: 10.3892/Or.2014.3537. Epub 2014 Oct 10. Cd38 Is Highly Expressed And Affects The Pi3K/Akt Signaling Pathway In Cervical Cancer.
Xiao S, Zhou Y, Yi W, Luo G, Jiang B, Tian Q, Li Y, Xue M. Int J Oncol. 2015 Apr;46(4):1677-84. Doi: 10.3892/Ijo.2015.2873. Epub 2015 Feb 4. Fra-1 Is Downregulated In Cervical Cancer Tissues And Promotes Cervical Cancer Cell Apoptosis By P53 Signa...

Customer Q&As

Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WB
A: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
A: You can find the immunogen sequence under "Immunogen" and clonality in the product name.
Q: What is the expected band size? Why is it different than the observed band size?
A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands

3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.

4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
A: Check our protocols under the tech support tab.
Q: What are some alternative names that could be used to describe this product?
A: Some common names include but are not limited to mdm2 antibody, hdm2 antibody, mtbp antibody