Recombinant Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20662-29] to Smad1 (phospho S463 + S465)
- Suitable for: WB, Dot blot, IP
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29]
See all Smad1 primary antibodies -
Description
Rabbit monoclonal [EPR20662-29] to Smad1 (phospho S463 + S465) -
Host species
Rabbit -
Specificity
Based on sequence homology this antibody also reacts with Smad5 (phospho S463/S465) and Smad9 (phospho S465/S467). -
Tested applications
Suitable for: WB, Dot blot, IPmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Dot Blot: Smad1 (phospho S463/S465) peptide, Smad1 (phospho S465) peptide, Smad5 (phospho S463/S465) peptide, Smad5 (phospho S465) peptide. WB: Calyculin (ab141784) treated HeLa cell lysate; BMP2 treated NIH/3T3 cell lysate. IP: BMP2 treated NIH/3T3 cell lysate.
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General notes
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20662-29 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab214423 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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WB |
1/1000. Detects a band of approximately 60 kDa (predicted molecular weight: 52 kDa).
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Dot blot |
1/1000.
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IP |
1/30.
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Notes |
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WB
1/1000. Detects a band of approximately 60 kDa (predicted molecular weight: 52 kDa). |
Dot blot
1/1000. |
IP
1/30. |
Target
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Function
Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD1 is a receptor-regulated SMAD (R-SMAD). SMAD1/OAZ1/PSMB4 complex mediates the degradation of the CREBBP/EP300 repressor SNIP1. -
Tissue specificity
Ubiquitous. Highest expression seen in the heart and skeletal muscle. -
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 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. Co-localizes with LEMD3 at the nucleus inner membrane. - Information by UniProt
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Database links
- Entrez Gene: 4086 Human
- Entrez Gene: 17125 Mouse
- Omim: 601595 Human
- SwissProt: Q15797 Human
- SwissProt: P70340 Mouse
- Unigene: 604588 Human
- Unigene: 223717 Mouse
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Alternative names
- BSP-1 antibody
- BSP1 antibody
- HsMAD1 antibody
see all
Images
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All lanes : Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) grown in serum-free media overnight, whole cell lysate
Lane 2 : HeLa grown in serum-free media overnight, then treated with 100ng/ml Calyculin A (ab141784) for 15 minutes, Calyculin A was removed, followed by treatment with 100ng/ml BMP2 for 30 minutes, whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) grown in serum-free media overnight, whole cell lysate
Lane 4 : NIH/3T3 grown in serum-free media overnight, then treated with 50ng/ml BMP2 for 30 minutes, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking/Dilution: 5% NFDM/TBST.
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Dot blot analysis of Smad1 (phospho S463/S465) peptide (Lane 1), Smad1 (phospho S463) peptide (Lane 2), Smad1 (phospho S465) peptide (Lane 3), Smad1 non-phospho peptide (Lane 4), Smad5 (phospho S463/S465) peptide (Lane 5), Smad5 (phospho S463) peptide (Lane 6), Smad5 (phospho S465) peptide (Lane 7) and Smad5 non-phospho peptide (Lane 8) using ab214423 at 1/1,000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.
Blocking and Diluting buffer and concentration: 5% NFDM /TBST.
Exposure time: 30 seconds.
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Smad1 (phospho S463/S465) was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate with ab214423 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214423 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution
Lane 1: NIH/3T3 (mouse embryonic fibroblast) grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate 10 μg (Input).
Lane 2: ab214423 IP in NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214423 in NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysateBlocking and dilution buffer: 5% NFDM/TBST.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab214423 has been referenced in 3 publications.
- Kato T et al. Candesartan prevents arteriopathy progression in cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy model. J Clin Invest 131:N/A (2021). PubMed: 34779414
- Kim MS & Jin W TrkB Inhibits the BMP Signaling-Mediated Growth Inhibition of Cancer Cells. Cancers (Basel) 12:N/A (2020). PubMed: 32731498
- Pei DD et al. Contribution of Mitophagy to Cell-Mediated Mineralization: Revisiting a 50-Year-Old Conundrum. Adv Sci (Weinh) 5:1800873 (2018). PubMed: 30356983