Anti-MDMX/MDM4 antibody (ab16058)
Key features and details
- Rabbit polyclonal to MDMX/MDM4
- Suitable for: ICC/IF, WB
- Reacts with: Cow, Human
- Isotype: IgG
Overview
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Product name
Anti-MDMX/MDM4 antibody
See all MDMX/MDM4 primary antibodies -
Description
Rabbit polyclonal to MDMX/MDM4 -
Host species
Rabbit -
Specificity
Two bands were detected with the antibody in HeLa nuclear extract, at ~80 and 100 kDa. The 80 kDa band was no longer detected after the addition of competing peptide, suggesting that this band corresponds to MDMX/MDM4. MDMX/MDM4 has been shown previously to migrate aberrantly at an ~80 kDa size (see deGraaf, et.al., J Biol Chem. 2003 Oct 3;278(40):38315-24). The 100 kDa band is not competed away by the addition of the immunizing peptide, suggesting that it is nonspecific cross-reactivity.
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Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
This product was produced with the following immunogens:
Synthetic peptide corresponding to Human MDMX/MDM4 aa 200-300 (internal sequence) conjugated to keyhole limpet haemocyanin.
(Peptide available asab16383)
Synthetic peptide corresponding to Human MDMX/MDM4 aa 100-200 (internal sequence) conjugated to keyhole limpet haemocyanin.
(Peptide available asab16383) -
Positive control
- Recombinant Human MDMX/MDM4 protein (ab114467) can be used as a positive control in WB. This antibody gave a positive signal in the following nuclear cell extract: HeLa. This antibody gave a positive signal in the following Methanol fixed cell lines: HeLa.
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General notes
This product was previously labelled as MDMX
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab16058 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB HumanApplication Abreviews Notes ICC/IF (1) WB Notes Application notesICC/IF: Use at a concentration of 5 µg/ml.
WB: 1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 58 kDa). Can be blocked with MDMX peptide.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.Target
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Function
Inhibits p53/TP53- and TP73/p73-mediated cell cycle arrest and apoptosis by binding its transcriptional activation domain. Inhibits degradation of MDM2. Can reverse MDM2-targeted degradation of TP53 while maintaining suppression of TP53 transactivation and apoptotic functions. -
Tissue specificity
Expressed in all tissues tested with high levels in thymus. -
Sequence similarities
Belongs to the MDM2/MDM4 family.
Contains 1 RanBP2-type zinc finger.
Contains 1 RING-type zinc finger.
Contains 1 SWIB domain. -
Domain
Region I is sufficient for binding TP53 and inhibiting its G1 arrest and apoptosis functions. It also binds TP73. Region II contains most of a central acidic region and a putative C4-type zinc finger. The RING finger domain which coordinates two molecules of zinc mediates the heterooligomerization with MDM2. -
Post-translational
modificationsUbiquitinated. Deubiquitinated by USP2; leading to stabilize it. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 4194 Human
- Omim: 602704 Human
- SwissProt: O15151 Human
- Unigene: 497492 Human
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Alternative names
- DKFZp781B1423 antibody
- Double minute 4 antibody
- Double minute 4 human homolog of p53 binding protein antibody
see all
Images
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Western blot - Anti-MDMX/MDM4 antibody (ab16058)All lanes : Anti-MDMX/MDM4 antibody (ab16058) at 1/1000 dilution
Lane 1 : 20ug calf thymus histone
Lane 2 : 20ug HeLa nuclear extract
Lane 3 : 20ug calf thymus histone with Human MDMX/MDM4 peptide (ab16383) at 1 µg/ml
Lane 4 : 20ug HeLa nuclear extract with Human MDMX/MDM4 peptide (ab16383) at 1 µg/ml
Secondary
All lanes : Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa (possible cross reactivity)Two bands were detected with the antibody in HeLa nuclear extract, at ~80 and 100 kDa. The 80 kDa band was no longer detected after the addition of competing peptide, suggesting that this band corresponds to MDMX/MDM4. The 100 kDa band is not competed away by the addition of the immunizing peptide, suggesting that it is nonspecific cross-reactivity.
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Immunocytochemistry/ Immunofluorescence - Anti-MDMX/MDM4 antibody (ab16058)ICC/IF image of ab16058 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab16058, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
Protocols
References (7)
ab16058 has been referenced in 7 publications.
- Fan Y et al. Recombinant Dual-target MDM2/MDMX Inhibitor Reverses Doxorubicin Resistance through Activation of the TAB1/TAK1/p38 MAPK Pathway in Wild-type p53 Multidrug-resistant Breast Cancer Cells. J Cancer 11:25-40 (2020). PubMed: 31892970
- Fan Y et al. Dual-target MDM2/MDMX inhibitor increases the sensitization of doxorubicin and inhibits migration and invasion abilities of triple-negative breast cancer cells through activation of TAB1/TAK1/p38 MAPK pathway. Cancer Biol Ther 20:617-632 (2019). PubMed: 30462562
- Zu Y et al. Tan IIA inhibits H1299 cell viability through the MDM4-IAP3 signaling pathway. Mol Med Rep 17:2384-2392 (2018). PubMed: 29207086
- Xu XL et al. CPUY201112, a novel synthetic small-molecule compound and inhibitor of heat shock protein Hsp90, induces p53-mediated apoptosis in MCF-7 cells. Sci Rep 6:19004 (2016). PubMed: 26743233
- Zhuang C et al. A novel drug discovery strategy: mechanistic investigation of an enantiomeric antitumor agent targeting dual p53 and NF-?B pathways. Oncotarget 5:10830-9 (2014). WB . PubMed: 25350970
- Liu Z et al. A Fluorescent Probe for Imaging p53-MDM2 Protein-Protein Interaction. Chem Biol Drug Des N/A:N/A (2014). WB ; Human . PubMed: 25226874
- Tabe Y et al. MDM2 antagonist nutlin-3 displays antiproliferative and proapoptotic activity in mantle cell lymphoma. Clin Cancer Res 15:933-42 (2009). PubMed: 19188164
Images
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Western blot - Anti-MDMX/MDM4 antibody (ab16058)All lanes : Anti-MDMX/MDM4 antibody (ab16058) at 1/1000 dilution
Lane 1 : 20ug calf thymus histone
Lane 2 : 20ug HeLa nuclear extract
Lane 3 : 20ug calf thymus histone with Human MDMX/MDM4 peptide (ab16383) at 1 µg/ml
Lane 4 : 20ug HeLa nuclear extract with Human MDMX/MDM4 peptide (ab16383) at 1 µg/ml
Secondary
All lanes : Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa (possible cross reactivity)Two bands were detected with the antibody in HeLa nuclear extract, at ~80 and 100 kDa. The 80 kDa band was no longer detected after the addition of competing peptide, suggesting that this band corresponds to MDMX/MDM4. The 100 kDa band is not competed away by the addition of the immunizing peptide, suggesting that it is nonspecific cross-reactivity.
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Immunocytochemistry/ Immunofluorescence - Anti-MDMX/MDM4 antibody (ab16058)
ICC/IF image of ab16058 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab16058, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
