Overview

  • Product name

    Mouse sTNF RI ELISA Kit (TNFRSF1A)
    See all TNF Receptor I kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 5 1.5%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 13.8%
  • Sample type

    Cell culture supernatant, Urine, Serum, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    1.2 pg/ml
  • Range

    7.81 pg/ml - 500 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 103 101% - 109%
    Serum 113 107% - 118%
    Cell culture media 95 94% - 95%
    EDTA Plasma 102 97% - 109%
    Citrate Plasma 106 97% - 112%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Rabbit, Goat, Horse, Cow, Dog
  • Product overview

    Abcam’s sTNF RI (TNFRSF1A) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of sTNF RI protein in mouse serum, plasma, urine and cell culture supernatant.

    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing 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.

  • Notes

    Tumor necrosis factor receptor superfamily member 1A (TNF RI) is a transmembrane protein with an extracellular domain that binds to TNF alpha. This extracellular domain can be proteolytically cleaved to make soluble TNF RI. The standard protein in this kit is soluble TNF RI and the capture and detector antibodies were raised against this region of TNF RI.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab202408 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • 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.

     

  • Standard curve comparison between mouse TNFRSF1A SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of sTNF RI were measured in duplicate and interpolated from the sTNF RI standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean sTNF RI concentration was determined to be 3,427 pg/mL in mouse serum, 2,484 pg/mL in mouse plasma (Citrate), and 2,982 pg/mL in mouse plasma (EDTA).

  • The concentrations of sTNF RI were measured in duplicate and interpolated from the sTNF RI standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean sTNF RI concentration was determined to be 1,754 pg/mL in mouse urine, 538.3 pg/mL L929 stimulated cell culture supernatant.

  • L929 and J774A.1 cells were grown in the absence (unstimulated) or presence of Phorbol Myristate Acetate (PMA) and phytohemagglutinin (PHA) (stimulated) for 3 days. sTNF RI was measured in 2-fold diluted cell culture supernatants of unstimulated and PMA/PHA stimulated L929 and J774A.1 and cell culture media. Measured values were interpolated from the sTNF RI Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed. sTNF RI is undetectable in media.

Protocols

References

ab202408 has not yet been referenced specifically in any publications.

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