SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit is a single-wash 90-min Simplestep used to quantify SMAD1 (pS463/S465 + Total SMAD1) with a sensitivity of 20 µg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
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- Nature metabolism 2:514-5312020Bone morphogenetic protein 8B promotes the progression of non-alcoholic steatohepatitis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMichele Vacca,Jack Leslie,Samuel Virtue,Brian Y H Lam,Olivier Govaere,Dina Tiniakos,Sophie Snow,Susan Davies,Kasparas Petkevicius,Zhen Tong,Vivian Peirce,Mette Juul Nielsen,Zsuzsanna Ament,Wei Li,Tomasz Kostrzewski,Diana Julie Leeming,Vlad Ratziu,Michael E D Allison,Quentin M Anstee,Julian L Griffin,Fiona Oakley,Antonio Vidal-PuigPubMed 32694734
Key facts
- Detection method
- Colorimetric
- Sample types
- Tissue Homogenate, Cell Lysate
- Assay type
- Semi-quantitative
- Reactive species
- Mouse, Human
- Range
- 10 - 1000 µg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 20 µg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
Target data
Function
Transcriptional modulator that plays a role in various cellular processes, including embryonic development, cell differentiation, and tissue homeostasis (PubMed:9335504). Upon BMP ligand binding to their receptors at the cell surface, is phosphorylated by activated type I BMP receptors (BMPRIs) and associates with SMAD4 to form a heteromeric complex which translocates into the nucleus acting as transcription factor (PubMed:33667543). In turn, the hetero-trimeric complex recognizes cis-regulatory elements containing Smad Binding Elements (SBEs) to modulate the outcome of the signaling network (PubMed:33667543). SMAD1/OAZ1/PSMB4 complex mediates the degradation of the CREBBP/EP300 repressor SNIP1. Positively regulates BMP4-induced expression of odontogenic development regulator MSX1 following IPO7-mediated nuclear import (By similarity).
Alternative names
BSP1, MADH1, MADR1, SMAD1, Mothers against decapentaplegic homolog 1, MAD homolog 1, Mothers against DPP homolog 1, JV4-1, Mad-related protein 1, SMAD family member 1, Transforming growth factor-beta-signaling protein 1, SMAD 1, Smad1, hSMAD1, BSP-1
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SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit is a single-wash 90-min Simplestep used to quantify SMAD1 (pS463/S465 + Total SMAD1) with a sensitivity of 20 µg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
Key facts
- Detection method
- Colorimetric
- Sample types
- Tissue Homogenate, Cell Lysate
- Assay type
- Semi-quantitative
- Reactive species
- Mouse, Human
- Range
- 10 - 1000 µg/mL
- Assay time
- 1h 30m
- Assay Platform
- Microplate
- Sensitivity
- = 20 µg/mL
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Overall | n 6 | C.V. 5.3 |
Sample pS463/S465 | n 6 | C.V. 2.3 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample Overall | n 3 | C.V. 5.6 |
Sample pS463/S465 | n 3 | C.V. 7.5 |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
SMAD1 (pS463/S465) and SMAD1 (Total) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of SMAD (pS463/S465) and Total SMAD1 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.
ASSAY SPECIFICITY The SMAD1 (pS463/S465) assay detects endogenous levels of SMAD1 (GenBank Accession NP_001003688) in cellular lysates, only when phosphorylated at Ser463/465. Based on sequence similarity, cross reaction to SMAD5 and SMAD8 may occur.
The SMAD1 (Total) assay detects endogenous levels of SMAD1 (GenBank Accession NP_001003688) in cellular lysates. Total SMAD1 assay kits detect SMAD1, irrespective of phosphorylation status. Based on sequence similarity, cross reaction to SMAD5 and SMAD8 may occur.
SPECIES REACTIVITY This kit detects SMAD1 (pS463/S465) and SMAD1 (Total) in Human and mouse cell culture extracts. Detection in rat samples is also expected. Other species should be tested on a case-by-case basis.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Smad1 also known as Mothers Against Decapentaplegic Homolog 1 is a protein that functions as an intracellular messenger. Its molecular weight is approximately 60 kDa. Smad1 is expressed in various tissues with higher levels found in bone and cartilage tissues. Mechanically Smad1 transmits signals from the cell membrane to the nucleus. It gets activated upon phosphorylation by receptor-regulated Smad kinases specifically. Once phosphorylated Smad1 forms a complex with Smad4 which enables its movement into the nucleus to regulate gene expression.
- Biological function summary
Smad1 plays a central role in bone morphogenetic protein (BMP) signaling pathways. It serves as part of a multiprotein complex allowing it to exert its functions effectively. This protein impacts cellular processes such as proliferation differentiation and apoptosis. It is very important in the development of tissues that require BMP signals. Through its ability to regulate gene expression depending on external BMP signals Smad1 contributes to the modulation of embryogenesis and tissue homeostasis.
- Pathways
Smad1 participates in the transforming growth factor-beta (TGF-beta) signaling pathway and the BMP signaling pathway. The BMP pathway significantly influences bone formation and maintenance. Within this network Smad1 closely interacts with other proteins like BMP receptors and Smad4. In the context of TGF-beta signaling Smad1 aligns functionally alongside Smad2 and Smad3 although typically it is more BMP-specific. The interconnectedness with these pathways highlights Smad1's importance in cellular responses to growth factors.
- Associated diseases and disorders
Smad1 has connections to various conditions notably cancer and fibrodysplasia ossificans progressiva (FOP). Alterations or mutations in Smad1 signaling can lead to uncontrolled cell growth and cancer development due to the disruption of normal cell cycle control. In FOP aberrant signaling involving Smad1 leads to the formation of bone in soft tissues. This disorder also involves protein interactions with ACVR1 a type I BMP receptor which drives the pathological ossification process.
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6 product images
Sandwich ELISA - SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit (ab186035)
Typical cell lysate dilution series.
Example of a typical SMAD1 (pS463/S465) and SMAD1 (Total) cell lysate dilution series. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit (ab186035)
Example of a typical SMAD1 (pS463/S465) and SMAD1 (Total) cell lysate dilution series in 1X Cell Extraction Buffer PTR. The SMAD1 lysate dilution series was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit (ab186035)
Linearity 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.
Sandwich ELISA - SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit (ab186035)
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).
Sandwich ELISA - SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit (ab186035)
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.
Sandwich ELISA - SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit (ab186035)
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.
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