Recombinant Anti-SMAD5 (phospho S463 + S465) antibody [EP728(2)AY] (ab76296)
![Western blot - Anti-SMAD5 (phospho S463 + S465) antibody [EP728(2)AY] (ab76296)](https://www.abcam.com/ps/products/76/ab76296/Images/ab76296-91671-anti-smad5-phospho-s463-s465-antibody-ep7282ay-western-blot.jpg "Western blot - Anti-SMAD5 (phospho S463 + S465) antibody [EP728(2)AY] (ab76296)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP728(2)AY] to SMAD5 (phospho S463 + S465)
- Suitable for: WB, ICC/IF
- Reacts with: Human
Overview
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Product name
Anti-SMAD5 (phospho S463 + S465) antibody [EP728(2)AY]
See all SMAD5 primary antibodies -
Description
Rabbit monoclonal [EP728(2)AY] to SMAD5 (phospho S463 + S465) -
Host species
Rabbit -
Specificity
Detects SMAD5 phosphorylated at Serine 463 and 465. This antibody may cross-react with Smad1 Phospho (pS463/465) and Smad9 Phospho (pS465/467). -
Tested applications
Suitable for: WB, ICC/IFmore details
Unsuitable for: Flow Cyt,IHC-P or IP -
Species reactivity
Reacts with: Human -
Immunogen
A phospho specific peptide corresponding to residues surrounding Serine 463 and 465 of human SMAD5.
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Positive control
- HeLa cytoplasmic or nuclear lysate treated with BMP-4 HeLa cells
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General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EP728(2)AY -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab76296 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 |
|---|---|---|
| WB | 1/500. Predicted molecular weight: 52 kDa. | |
| ICC/IF | 1/50 - 1/100. |
Target
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Function
Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD5 is a receptor-regulated SMAD (R-SMAD). -
Tissue specificity
Ubiquitous. -
Sequence similarities
Belongs to the dwarfin/SMAD family.
Contains 1 MH1 (MAD homology 1) domain.
Contains 1 MH2 (MAD homology 2) domain. -
Post-translational
modificationsPhosphorylated on serine by BMP (bone morphogenetic proteins) type 1 receptor kinase.
Ubiquitin-mediated proteolysis by SMAD-specific E3 ubiquitin ligase SMURF1. -
Cellular localization
Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4. - Information by UniProt
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Database links
- Entrez Gene: 4090 Human
- Omim: 603110 Human
- SwissProt: Q99717 Human
- Unigene: 167700 Human
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Alternative names
- DKFZp781C1895 antibody
- DKFZp781O1323 antibody
- Dwfc antibody
see all
Images
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Western blot - Anti-SMAD5 (phospho S463 + S465) antibody [EP728(2)AY] (ab76296)All lanes : Anti-SMAD5 (phospho S463 + S465) antibody [EP728(2)AY] (ab76296) at 1/500 dilution
Lane 1 : HeLa cell lysate - untreated
Lane 2 : HeLa cell lysate - treated with BMP-4
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 52 kDa
Observed band size: 52 kDa -
![Immunocytochemistry/ Immunofluorescence - Anti-SMAD5 (phospho S463 + S465) antibody [EP728(2)AY] (ab76296)](https://www.abcam.com/ps/products/76/ab76296/Images/ab76296-91670-anti-smad5-phospho-s463-s465-antibody-ep7282ay-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-SMAD5 (phospho S463 + S465) antibody [EP728(2)AY] (ab76296)Immunofluorescent staining of HeLa cells using ab76296 at a 1/50-1/100 dilution. -
![Anti-SMAD5 (phospho S463 + S465) antibody [EP728(2)AY] (ab76296)](https://www.abcam.com/ps/products/76/ab76296/Images/ab76296-3-benefits-of-recombinant-antibodies.png)
Anti-SMAD5 (phospho S463 + S465) antibody [EP728(2)AY] (ab76296)
Protocols
Datasheets and documents
References (4)
ab76296 has been referenced in 4 publications.
- Zhou Q et al. A New Insight of Kartogenin Induced the Mesenchymal Stem Cells (MSCs) Selectively Differentiate into Chondrocytes by Activating the Bone Morphogenetic Protein 7 (BMP-7)/Smad5 Pathway. Med Sci Monit 25:4960-4967 (2019). PubMed: 31271564
- Cui Y et al. Single-Cell Transcriptome Analysis Maps the Developmental Track of the Human Heart. Cell Rep 26:1934-1950.e5 (2019). PubMed: 30759401
- Ghiabi P et al. Breast cancer cells promote a notch-dependent mesenchymal phenotype in endothelial cells participating to a pro-tumoral niche. J Transl Med 13:27 (2015). WB ; Human . PubMed: 25623554
- Nili M et al. Soluble Repulsive Guidance Molecule c/Hemojuvelin Is a Broad Spectrum Bone Morphogenetic Protein (BMP) Antagonist and Inhibits both BMP2- and BMP6-mediated Signaling and Gene Expression. J Biol Chem 285:24783-92 (2010). WB ; Human . PubMed: 20530805
