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
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Cow, Human -
Immunogen
Synthetic peptide corresponding to Human MDMX/MDM4 aa 200-300 (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. ICC/IF: HeLa cells
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research 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.
Application | Abreviews | Notes |
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ICC/IF | (1) |
Use a concentration of 5 µg/ml.
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WB |
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 58 kDa).
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Notes |
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ICC/IF
Use a concentration of 5 µg/ml. |
WB
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 58 kDa). |
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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ab16058 staining MDMX/MDM4 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab16058 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab16058 has been referenced in 10 publications.
- Uddin MA et al. Hsp90 inhibition protects brain endothelial cells against LPS-induced injury. Biofactors 48:926-933 (2022). PubMed: 35266593
- Elmarakeby HA et al. Biologically informed deep neural network for prostate cancer discovery. Nature 598:348-352 (2021). PubMed: 34552244
- Sahin I et al. Hyperprogression of a mismatch repair-deficient colon cancer in a humanized mouse model following administration of immune checkpoint inhibitor pembrolizumab. Oncotarget 12:2131-2146 (2021). PubMed: 34676046
- 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