SMAD1 (pS463/465) ELISA Kit is a single-wash 90-min Simplestep used to quantify SMAD1 (pS463/S465) with a sensitivity of 10 µg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Semi-quantitative sandwich ELISA - 450 nm readout : works on any standard plate reader
Application | Reactivity | Dilution info | Notes |
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Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
SMAD1 (pS463/465) ELISA Kit is a single-wash 90-min Simplestep used to quantify SMAD1 (pS463/S465) with a sensitivity of 10 µg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Semi-quantitative sandwich ELISA - 450 nm readout : works on any standard plate reader
Sample | n | C.V. |
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Sample C2C12 | n 6 | C.V. 2.3 |
Sample | n | C.V. |
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Sample C2C12 | n 3 | C.V. 7.5 |
SMAD1 (pS463/465) ELISA Kit ab186036 is a rapid single-wash 90-min sandwich ELISA kit to measure SMAD1 (pS463/465) in Cell Lysates and Tissue Homogenates. This Simplestep sensitivity is 10 µg/mL.
How the assay works
SMAD1 (pS463/465) SimpleStep ELISA®employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details.
SMAD1 (pS463/465) ELISA Kit ab186036 protocol summary
1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm
How other researchers are using SMAD1 (pS463/465) ELISA Kit ab186036
SMAD1 (pS463/465) ELISA Kit ab186036 has been used to measure BMP signaling in heterotopic ossification(1).
References:
(1) L. Li, et.al., 2019, PMID: 31508317
Related and Recommended products
SMAD1 (pS463/465) ELISA Kit ab186036 is used to measure SMAD1 phosphorylation in the BMP signaling pathway.
Other ELISA kits to measure the BMP signaling pathway include:
- SMAD3 (pS423/S425) ELISA Kit SMAD3 (pS423/S425) ELISA Kit ab186038
- Human SMAD4 ELISA Kit Human SMAD4 ELISA Kit ab253211
- Human BMP-4 ELISA Kit Human BMP-4 ELISA Kit ab231930
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Smad1 also known as Mothers against decapentaplegic homolog 1 (MADH1) is a protein encoded by the SMAD1 gene in humans. It has a molecular mass of approximately 53 kDa and functions as a receptor-regulated Smad protein. Smad1 primarily transmits signals from the bone morphogenetic proteins (BMPs) a group of growth factors and cytokines. Expression of Smad1 occurs in various tissues including the lung heart and kidney. It plays an important role in mediating signaling pathways within different cellular environments.
Smad1 participates in the transmission of BMP signals from the cell surface to the nucleus ensuring transcriptional regulation of target genes. In the presence of BMPs Smad1 forms complexes with Smad4 upon activation. These Smad complexes then translocate to the nucleus where they regulate gene expression. Smad1 influences cellular responses such as proliferation differentiation and apoptosis. In particular it plays a significant role in bone development and osteogenesis.
Smad1 plays an important role within the BMP signaling pathway which is important for early development and tissue homeostasis. Within this pathway BMPs trigger the phosphorylation of Smad1 which then associates with Smad4 to propagate downstream signaling. Another associated pathway includes the TGF-beta signaling pathway where Smads like Smad2 and Smad3 show functional similarities and differences with Smad1. The interactions of Smad1 with related proteins like Smad4 highlight its significant contribution to cellular processes regulated by these pathways.
Smad1 associates with various pathological conditions. For instance aberrant Smad1 signaling has implications in cancer particularly in processes like tumor progression and metastasis. Disruptions in the BMP pathway involving Smad1 and Smad4 can influence the onset and development of disorders such as pulmonary hypertension where inappropriate cell growth and remodeling occur. Understanding Smad1's role in these contexts provides insights into potential therapeutic avenues for treatment and management of these conditions.
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Typical cell lysate dilution series.
Example 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.
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).
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.
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.
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.
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