Recombinant Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free (ab228457)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20662-20] to Smad1 (phospho S463 + S465) - BSA and Azide free
- Suitable for: IP, WB, ICC/IF, Dot blot
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free
See all Smad1 primary antibodies -
Description
Rabbit monoclonal [EPR20662-20] to Smad1 (phospho S463 + S465) - BSA and Azide free -
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: IP, WB, ICC/IF, Dot blotmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- ICC/IF: NIH3T3 cells FBS-deprived overnight before treatment with 50 ng/ml hBMP2 for 30min.
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General notes
ab228457 is the carrier-free version of ab226821.
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
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20662-20 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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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 ab228457 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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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 60 kDa (predicted molecular weight: 52 kDa).
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ICC/IF |
Use at an assay dependent concentration.
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Dot blot |
Use at an assay dependent concentration.
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Notes |
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IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 60 kDa (predicted molecular weight: 52 kDa). |
ICC/IF
Use at an assay dependent concentration. |
Dot blot
Use at an assay dependent concentration. |
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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Dot blot analysis of Smad1 (phospho S463 + S465) labeled with ab226821 at 1/1000 dilution.
Lane 1: Smad1 (phospho S463/S465) peptide;
Lane 2: Smad1 (phospho S463) peptide;
Lane 3: Smad1 (phospho S465) peptide;
Lane 4: Smad1 peptide (not phosphorylated);
Lane 5: Smad5 (phospho S463/S465) peptide;
Lane 6: Smad5 (phospho S463) peptide;
Lane 7: Smad5 (phospho S465) peptide;
Lane 9: Smad5 peptide (not phosphorylated).
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Based on sequence homology, this antibody cross-reacts with Smad5 (phospho S463/S465) and Smad9 (phospho S465/S467).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab226821).
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Smad 1 (phospho S463 + S465) was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryo fibroblast cell line) grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate with ab226821 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab226821 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate 10 μg (Input).
Lane 2: ab226821 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 ab226821 in NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab226821).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling Smad1 (phospho S463 + S465) with ab226821 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining in hBMP2-treated NIH/3T3 cells. Cells were FBS-deprived overnight before treatment with 50 ng/ml hBMP2 for 30 minutes.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab226821).
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)
ab228457 has not yet been referenced specifically in any publications.