Overview

  • Product name
    Human Interferon gamma ELISPOT Kit
    See all Interferon gamma kits
  • Detection method
    Colorimetric
  • Sample type
    Cell culture supernatant, Cell culture extracts
  • Assay type
    Sandwich (qualitative)
  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Human, Monkey
  • Product overview

    The Elispot assay is designed to enumerate cytokine producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in-vitro manipulations allowing cytokine production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation, and the follow-up of such frequency during a treatment and/or a pathological state. Elispot assay constitutes an ideal tool in the TH1 / TH2 response, vaccine development, viral infection monitoring and treatment, cancerology, infectious diseases, autoimmune diseases and transplantation.

    This Elispot assay is based on sandwich immuno-enzyme technology. Cell secreted cytokines or soluble molecules are captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured cytokines are revealed by tracer antibodies and appropriate conjugates.

    Principal:

    After cell stimulation, locally produced cytokines are captured by a specific monoclonal antibody. After cell lysis, trapped cytokine molecules are revealed by a secondary biotinylated detection antibody, which is in turn recognised by streptavidin conjugated to alkaline phosphatase. PVDF-bottomed-well plates are then incubated with BCIP/NBT substrate. Colored "purple" spots indicate cytokine production by individual cells.

     

    Recognizes natural human Interferon Gamma

  • Tested applications
    Suitable for: ELISpotmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 5 x 96 tests 10 x 96 tests
    96 PVDF-bottomed-well plates. 5 units 10 units
    Bovine Serum Albumin 1 x 1g 2 x 1g
    Human IFN Gamma  Capture antibody 1 x 0.5ml 2 x 0.5ml
    Biotinylated Human IFNγ Detection Antibody 1 x 0.55ml 2 x 0.55ml
    Ready-to-use BCIP/NBT substrate buffer 1 x 50ml 2 x 50ml
    Streptavidin - Alkaline Phosphatase conjugated 1 x 50µl 2 x 50µl
  • 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
    • IFG
    • IFI
    • IFN gamma
    • IFN, immune
    • IFN-gamma
    • IFNG
    • IFNG_HUMAN
    • Immune interferon
    • Interferon gamma
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab46569 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
ELISpot Use at an assay dependent dilution.

References

This product has been referenced in:
  • Saron WAA  et al. Flavivirus serocomplex cross-reactive immunity is protective by activating heterologous memory CD4 T cells. Sci Adv 4:eaar4297 (2018). Read more (PubMed: 29978039) »
See 1 Publication for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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