Overview

  • Product name

    Mouse sTNF RII ELISA Kit (TNFRSF1B)
    See all Soluble TNF Receptor II kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 5 5.5%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 8.6%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    1 pg/ml
  • Range

    6.25 pg/ml - 400 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 104 93% - 113%
    Serum 120 112% - 129%
    Cell culture media 100 97% - 106%
    EDTA Plasma 113 110% - 118%
    Citrate Plasma 114 100% - 125%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

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

    Abcam’s sTNF RII (TNFRSF1B) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of sTNF RII 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 1B (TNF RII) is a transmembrane protein with an extracellular domain that binds to TNF alpha. This extracellular domain can be proteolytically cleaved to make soluble TNF RII. The standard protein in this kit is soluble TNF RII and the capture and detector antibodies were raised against this region of TNF RII.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse sTNF RII Capture Antibody 1 x 600µl
    10X Mouse sTNF RII Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BC 1 x 6ml
    Mouse sTNF RII Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Relevance

    TNFRII (TNF Receptor II) is a member of the TNFR family of transmembrane proteins, and is expressed in immune cells and certain endothelial cells. It is a high affinity receptor for TNF alpha but manifests a lower affinity to TNF beta. Signaling through this receptor regulates various biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, coagulation, and neurotransmission. TNFRII consists of a 240 amino acid residue extracellular ligand binding portion, a 27 amino acid residue transmembrane segment and a 173 amino acid residue cytoplasmic domain. Soluble TNFRII (Soluble TNF Receptor II) is a 268 amino acid isoform containing the extracellular ligand-binding portion of TNFRII. It binds to TNF alpha and is capable of inhibiting TNF alpha induced activities by acting as a decoy receptor.
  • Alternative names

    • CD120b
    • Etanercept
    • p75
    • p80 TNF alpha receptor
    • TBP2
    • TBPII
    • TNFR2
    • TNFRII
    • Tumor necrosis factor binding protein 2
    • Tumor necrosis factor receptor 2
    • Tumor necrosis factor receptor superfamily member 1B
    • Tumor necrosis factor receptor superfamily member 1b, membrane form
    • Tumor necrosis factor receptor type II
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab202412 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 TNFRSF1B SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 4-fold increase in sensitivity.

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

  • The concentrations of sTNF RII were measured in duplicate and interpolated from the sTNF RII standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean sTNF RII concentration was determined to be 12 ng/mL in serum, 13.2 ng/mL in plasma (Citrate), and 12.8 ng/mL plasma (EDTA).

  • The concentrations of sTNF RII were measured in duplicate and interpolated from the sTNF RII standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean sTNF RII concentration was determined to be 13.3 ng/mL in urine and 3.9 ng/mL in RAW264.7 stimulated cell culture supernatant.

  • Cells were grown in the absence (unstimulated) or presence of Phorbol Myristate Acetate (PMA) and phytohemagglutinin (PHA) (stimulated) for 3 days. sTNF RII was measured in 2-fold diluted cell culture supernatants of unstimulated and PMA/PHA stimulated RAW264.7, L929, an J774A.1 and cell culture media. Measured values were interpolated from the sTNF RII Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed. sTNF RII was undetectable in media.

Protocols

References

ab202412 has not yet been referenced specifically in any publications.

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