SMAD1 (pS463/465) ELISA Kit (ab186036)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 10 µg/ml
- Range: 10 µg/ml - 500 µg/ml
- Sample type: Cell Lysate, Tissue Homogenate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Mouse, Human
Overview
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Product name
SMAD1 (pS463/465) ELISA Kit
See all Smad1 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% C2C12 6 2.3% Inter-assay Sample n Mean SD CV% C2C12 3 7.5% -
Sample type
Cell Lysate, Tissue Homogenate -
Assay type
Semi-quantitative -
Sensitivity
10 µg/ml -
Range
10 µg/ml - 500 µg/ml -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat
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Product overview
Abcam’s SMAD1 (pS463/S465) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of SMAD1 (pS463/S465) protein in human and mouse cells.
The SimpleStep ELISA™ employs a labeled capture and detector antibody which immunocaptures the sample analyte in solution. This entire complex (capture antibody/protein/detector antibody) is in turn immobilized in the well by immunoaffinity via the anti-tag antibody. Samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material; the TMB substrate is then added. The reaction is stopped by addition of Stop Solution which stops the color development and completes any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
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Notes
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Microplate
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Wash Buffer PT 1 x 15ml 50X Cell Extraction Enhancer Solution 1 x 1ml 5X Cell Extraction Buffer PTR 1 x 12ml Lyophilized SMAD1 (pS463/S465) Control Lysate 1 vial Plate Seal 1 unit SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit SMAD1 (pS463/S465) Capture Antibody 1 x 3ml SMAD1 (pS463/S465) Detector Antibody 1 x 3ml Stop Solution 1 x 12ml TMB Substrate 1 x 12ml -
Research areas
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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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Alternative names
- BSP-1
- BSP1
- HsMAD1
see all -
Database links
- Entrez Gene: 4086 Human
- Entrez Gene: 17125 Mouse
- Entrez Gene: 25671 Rat
- Omim: 601595 Human
- SwissProt: Q15797 Human
- SwissProt: P70340 Mouse
- SwissProt: P97588 Rat
- Unigene: 604588 Human
see all
Images
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Other - SMAD1 (pS463/465) ELISA Kit (ab186036)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Typical cell lysate dilution seriesExample of a typical SMAD1 (pS463/S465) cell lysate dilution series. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Linearity of dilutionLinearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1X Cell Extraction Buffer PTR. Data from duplicate measurements of SMAD1 (pS463/S465) are normalized and plotted.
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Comparison of total SMAD1 expression in different cell lines.Cell line analysis for Total SMAD1 from 300 µg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).
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SMAD1 (pS463/S465) phosphorylation in response to BMP-4 treatment.Induction of SMAD1 (pS463/S465) phosphorylation in HeLa cells in response to BMP-4 treatment. HeLa cells were cultured in 96-well tissue culture plates, serum-starved and treated (30 min) with a dose-range of BMP-4 before cell lysis. Data from quadruplicate measurements of SMAD1 (pS463/S465) are plotted.
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SMAD1 (pS463/S465) phosphorylation in response to dorsomorphin treatment.Inhibition of SMAD1 (pS463/S465) phosphorylation in HeLa cells in response to dorsomorphin treatment. HeLa cells were cultured in 96-well tissue culture plates and treated with a dose-range of dorsomorphin (30 min). Cells were then stimulated with BMP-4 (30 min) and lysed. Data from quadruplicate measurements of SMAD1 (pS463/S465) and SMAD1 (Total) are plotted.