Overview

  • Product name

    Mouse beta IG H3 ELISA Kit
    See all IGH3 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    EDTA plasma 8 4.4%
    Inter-assay
    Sample n Mean SD CV%
    EDTA plasma 3 4.9%
  • Sample type

    Cell culture supernatant, Cell culture extracts, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    40 pg/ml
  • Range

    0.16 ng/ml - 10 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture media 107 103% - 109%
    Heparin Plasma 107 105% - 110%
    EDTA Plasma 95 90% - 102%
    Citrate Plasma 89 83% - 93%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Human
  • Product overview

    Abcam’s beta IG H3 (TGFBI) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of beta IG H3 protein in mouse plasma, cell culture supernatants, cell and tissue extracts.

    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.

     

    Sensitivity: Samples diluted inSample Diluent NS: 70 pg/mL
                         Samples diluted in1X Cell Extraction Buffer PTR: 40 pg/mL

  • Notes

    Mouse beta IG H3 (TGFBI) is a 683 amino acid (aa) protein with a 23 aa signal peptide, one EMI domain, four FAS1 domains, and one RGD motif. Mouse beta IG H3 is expressed as a 75 kDa protein with no post-translational additions. Following secretion, cleavages at multiple positions near the C-terminal end release peptides with pro-apoptotic activity. The four FAS1 domains of Mouse beta IG H3 contain YH motifs, which allow Mouse beta IG H3 to bind to matrix fibronectin, collagen I, collagen VI, biglycan, and decorin. Mouse beta IG H3 is an adaptor protein that is induced in most cell types in response to TGF-beta stimulation. It is upregulated in keratinocytes and immature dendritic cells but downregulated in osteoblasts. Mouse beta IG H3 shares 91% aa sequence identity with human and porcine beta IG H3.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab206987 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.

     

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

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

  • Native mouse beta IG H3 was measured in 0.8% mouse plasma citrate, 0.8% mouse plasma EDTA, and 0.33% mouse plasma heparin samples diluted in a 2-fold dilution series in Sample Diluent NS. The concentrations of mouse beta IG H3 were measured in duplicate and interpolated from the mouse beta IG H3 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are graphed (mean +/- SD).

  • Recombinant mouse beta IG H3 was spiked into 50% cell culture media without FBS and into 10% cell culture media with FBS and diluted in a 2-fold dilution series in Sample Diluent NS. The concentrations of mouse beta IG H3 were measured in duplicate and interpolated from the mouse beta IG H3 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are graphed (mean +/- SD).

  • Native mouse beta IG H3 was measured in 100 μg/mL of mouse eyeball extract diluted in a 2-fold dilution series in Sample Diluent 1X Cell Extraction Buffer PTR. The concentrations of mouse beta IG H3 were measured in duplicate and interpolated from the mouse beta IG H3 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are graphed (mean +/- SD).

  • 3T3-L1 cells were cultured in cell culture media in the presence and absence 1 µM Dexamethasone, 0.5 mM IBMX, 1 µg/mL insulin for 10 days to generate adipocytes and undifferentiated cells respectively. At the end of the treatment, protein extraction was carried out according to section 11.4 of the booklet. Both 3T3-L1 adipocytes and undifferentiated cell extracts were measured in duplicate at 1 mg/mL using this kit. The concentrations of beta IG H3 were measured in duplicate and interpolated from the beta IG H3 standard. The interpolated dilution factor corrected values are graphed (mean +/- SD).

Protocols

References

ab206987 has not yet been referenced specifically in any publications.

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