Overview

  • Product name

    Human IFN gamma ELISA Kit, Fluorescent
    See all Interferon gamma kits
  • Detection method

    Fluorescent
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Supernatant 5 1.1%
    Inter-assay
    Sample n Mean SD CV%
    Supernatant 3 7.9%
  • Sample type

    Cell culture supernatant, Serum, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    25 pg/ml
  • Range

    0.06 ng/ml - 30 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 86 77% - 93%
    Serum 102 98% - 104%
    Hep Plasma 244 197% - 298%
    EDTA Plasma 122 117% - 124%
    Cit plasma 107 96% - 115%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Interferon-gamma (IFNG) in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Interferon-gamma (IFNG) protein in human serum, plasma, and cell culture supernatant samples.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint 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.  CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.


     

  • Notes

    IFN gamma is produced by lymphocytes activated by specific antigens or mitogens. IFN gamma, in addition to having antiviral activity, has important immunoregulatory functions. It is a potent activator of macrophages, it has antiproliferative effects on transformed cells and it can potentiate the antiviral and antitumor effects of the type I interferons.

     

  • 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
    100X Stoplight Red Substrate 1 x 120µl
    10X Human IFNG Capture Antibody 1 x 600µl
    10X Human IFNG Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Antibody Diluent CPI 1 x 6ml
    Human IFNG Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    Sample Diluent NBP 1 x 20ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 1 x 12ml
  • Research areas

  • Function

    Produced by lymphocytes activated by specific antigens or mitogens. IFN-gamma, in addition to having antiviral activity, has important immunoregulatory functions. It is a potent activator of macrophages, it has antiproliferative effects on transformed cells and it can potentiate the antiviral and antitumor effects of the type I interferons.
  • Tissue specificity

    Released primarily from activated T lymphocytes.
  • Involvement in disease

    In Caucasians, genetic variation in IFNG is associated with the risk of aplastic anemia (AA) [MIM:609135]. AA is a rare disease in which the reduction of the circulating blood cells results from damage to the stem cell pool in bone marrow. In most patients, the stem cell lesion is caused by an autoimmune attack. T-lymphocytes, activated by an endogenous or exogenous, and most often unknown antigenic stimulus, secrete cytokines, including IFN-gamma, which would in turn be able to suppress hematopoiesis.
  • Sequence similarities

    Belongs to the type II (or gamma) interferon family.
  • Post-translational
    modifications

    Proteolytic processing produces C-terminal heterogeneity, with proteins ending alternatively at Gly-150, Met-157 or Gly-161.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • IF 1
    • IFG
    • IFI
    • IFN gamma
    • IFN immune
    • IFN, immune
    • IFN-gamma
    • IFNG
    • IFNG_HUMAN
    • Immune interferon
    • Interferon gamma
    • Type II Interferon
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab229415 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.

     

  • The Interferon-gamma (IFNG) standard curve was prepared as described in Section 10.

  • PBMC were grown in the absence or presence of phytohemagglutinin (PHA) for 2 days. IFNG concentrations were measured in 12X and 6X diluted cell culture supernatants of the unstimulated PBMC and the stimulated PBMC, and media. Raw data values (mean +/-SD, n=3) are graphed. The dotted line represents zero sample background.

  • The concentrations of IFNG were interpolated from data values shown in Figure 3 using IFNG standard curve and corrected for sample dilution. The mean IFNG concentration was determined to be 1.8 ng/mL in unstimulated PBMC supernatants and 177.2 ng/mL in stimulated PBMC supernatants.

Protocols

References

ab229415 has not yet been referenced specifically in any publications.

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