Data & Images
|Product Name||Anti-MDM2/Hdm2 Antibody|
|Description||Rabbit IgG polyclonal antibody for E3 ubiquitin-protein ligase Mdm2(MDM2) detection. Tested with WB in Human.|
|Cite This Product||Anti-MDM2/Hdm2 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1378-1)|
|Replacement Item||This antibody may replace the following items: sc-56154|sc-56430|sc-965|sc-813|sc-7918|sc-5304|sc-812|sc-13161|sc-965-X from Santa Cruz Biotechnology.|
*This antibody is predicted to react with the above species based on antigen sequence similarities. Our Boster Guarantee covers the use of this product with the above species.
*Our Boster Guarantee covers the use of this product in the above tested applications.
**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.
|Recommended Detection Systems||Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2018!
|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.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Concentration||Add 0.2ml of distilled water will yield a concentration of 500ug/ml.|
|Storage||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.|
|Purification||Immunogen affinity purified.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Protein Name||E3 ubiquitin-protein ligase Mdm2|
|Molecular Weight||55233 MW|
|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. .|
|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.|
|Sequence Similarities||Belongs to the MDM2/MDM4 family.|
|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.|
|Alternative Names||E3 ubiquitin-protein ligase Mdm2;6.3.2.-;Double minute 2 protein;Hdm2;Oncoprotein Mdm2;p53-binding protein Mdm2;MDM2;|
|Research Areas|||cell biology|cell cycle|cell cycle inhibitors|p53| epigenetics and nuclear signaling|transcription|other factors| cancer|p53 pathway|oncoproteins/suppressors|oncoproteins|cell survival & death|ubiquitin & ubiquitin like modifiers|e3 ubiquitin ligases||
Background for E3 ubiquitin-protein ligase Mdm2
Dilution Ratios/Recommended Concentrations
At Boster we strive to provide the best Anti-MDM2/Hdm2 Antibody by testing all applications on non-spiked tissues and cell lines to ensure that the affinity of the antibody is enough to react to the endogenouse level of the target protein. Read more about our QC panel here.
|Recommended dilution ratios are listed below:|
Western blot, 0.1-0.5μg/ml, Human |
**Boster provides high sensitivity secondary antibody kits for Western blotting and IHC. For more info see Related Products below.
Anti-MDM2/Hdm2 Antibody Images
Click the images to enlarge.
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.
- Review by Stacy Wilson
"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. The bad: Having to resuspend and freeze in tiny aliquots was not my favorite part of using this. The bottom line: I would absolutely buy it again."
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.,