Astana Biomed Group, an authorized Abcam distributor in Central Asia

SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit (ab186035)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 20 µg/ml
  • Range: 10 µg/ml - 1000 µg/ml
  • Sample type: Cell Lysate, Tissue Homogenate
  • Detection method: Colorimetric
  • Assay type: Semi-quantitative
  • Reacts with: Mouse, Human

Overview

  • Product name

    SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit

  • Detection method

    Colorimetric

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    pS463/S465 6 2.3%
    Total 6 5.3%
    Inter-assay
    Sample n Mean SD CV%
    pS463/S465 3 7.5%
    Total 3 5.6%

  • Sample type

    Cell Lysate, Tissue Homogenate

  • Assay type

    Semi-quantitative

  • Sensitivity

    20 µg/ml

  • Range

    10 µg/ml - 1000 µg/ml

  • Assay time

    1h 30m

  • Assay duration

    One step assay

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat

  • Product overview

    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.

  • 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

Images

  • Other - SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit (ab186035)
    Other - SMAD1 (pS463/S465 + Total SMAD1) ELISA Kit (ab186035)

    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.

     

  • Typical cell lysate dilution series
    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.

  • Standard curve
    Standard curve

    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.

  • Linearity of dilution
    Linearity of dilution

    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.

  • Comparison of total SMAD1 expression in different cell lines.
    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.
    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.
    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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