Recombinant Anti-MDM2 (phospho S166) antibody [EPR1450(2)] - BSA and Azide free (ab249560)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1450(2)] to MDM2 (phospho S166) - BSA and Azide free
- Suitable for: Dot blot, IHC-P, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-MDM2 (phospho S166) antibody [EPR1450(2)] - BSA and Azide free
See all MDM2 primary antibodies -
Description
Rabbit monoclonal [EPR1450(2)] to MDM2 (phospho S166) - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Dot blot, IHC-P, WBmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Human
Predicted to work with: RatDoes not react with: Mouse -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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General notes
ab249560 is the carrier-free version of ab170880.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Affinity purified -
Clonality
Monoclonal -
Clone number
EPR1450(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab249560 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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Dot blot |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 55 kDa.
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Notes |
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Dot blot
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 55 kDa. |
Target
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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 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. -
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. -
Involvement in disease
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. -
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 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. -
Post-translational
modificationsPhosphorylated 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. -
Cellular 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. - Information by UniProt
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Database links
- Entrez Gene: 4193 Human
- Entrez Gene: 314856 Rat
- Omim: 164785 Human
- SwissProt: Q00987 Human
- Unigene: 484551 Human
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Alternative names
- ACTFS antibody
- Double minute 2 protein antibody
- E3 ubiquitin-protein ligase Mdm2 antibody
see all
Images
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All lanes : Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880) at 1/50000 dilution
Lane 1 : MCF7 cell lysate
Lane 2 : MCF7 cell lysate treated with IGF-1
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 55 kDaThis data was developed using ab170880, the same antibody clone in a different buffer formulation.
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This data was developed using ab170880, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labeling MDM2 (phospho S166) using ab170880 at a 1/50 dilution. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880) at 1/100000 dilution
Lane 1 : A549 (Human lung carcinoma epithelial cell) whole cell lysate
Lane 2 : A549 (Human lung carcinoma epithelial cell) treated with insulin at 1ug/ml for 150 minutes. Whole cell lysate
Lane 3 : A549 (Human lung carcinoma epithelial cell) treated with insulin at 1ug/ml for 150 minutes. Whole cell lysate. Then the membrane was incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 55 kDa
Observed band size: 90-140 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsThis data was developed using ab170880, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 2% BSA/TBST.
The molecular weight is the same with the one from this paper PMID: 25392082.
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This data was developed using ab170880, the same antibody clone in a different buffer formulation.Dot blot analysis of MDM2 (phospho S166) phospho peptide (Lane 1), MDM2 non-phospho peptide (Lane 2), labeling MDM2 (phospho S166) with ab170880 at a dilution of 1/1000. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/100000. Blocking and dilution buffer: 5% NFDM/TBST. Exposure time: 3 minutes.
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This data was developed using ab170880, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling MDM2 (phospho S166) using ab170880 at a 1/50 dilution. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab249560 has not yet been referenced specifically in any publications.