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Human Interferon Gamma + Granzyme B ELISPOT Set (ab48463)

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Key features and details

  • Assay type: Sandwich (qualitative)
  • Sample type: Suspension cells
  • Reacts with: Human

製品の概要

  • 製品名

    Human Interferon Gamma + Granzyme B ELISPOT Set

  • サンプルの種類

    Suspension cells

  • アッセイタイプ

    Sandwich (qualitative)

  • ステップ

    Multiple steps standard assay

  • 種交差性

    交差種: Human

  • 製品の概要

    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, oncology, 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.


    The dual colour ELISPOT allows you to monitor the production of two cytokines simultaneously in the same well.


    Principle of the Method


    After cell stimulation, locally produced cytokines are captured by IFN gamma and Granzyme B specific monoclonal antibodies. After cell lysis, trapped cytokine molecules are revealed by a secondary anti-IFN gamma FITC conjugated antibody and a biotinylated anti-Granzyme B antibody. Those are in turn recognised by anti-FITC HRP and streptavidin-AP conjugates. PVDF-bottomed-well plates are then incubated first with AEC substrate buffer, washed and subsequently incubated with BCIP/NBT. Coloured red/brownish spots indicate IFN gamma production while Granzyme B is revealed by blue/purple spots.

  • アプリケーション

    適用あり: ELISpotmore details

  • 試験プラットフォーム

    Reagents

製品の特性

  • 保存方法

    Store at +4°C. Please refer to protocols.
  • 内容 5 x 96 tests 10 x 96 tests 15 x 96 tests 20 x 96 tests
    10 x concentrate buffer for the preparation of AEC buffer 1 x 5ml 2 x 5ml 3 x 5ml 4 x 5ml
    50 x concentrate AEC substrate buffer 1 x 1ml 2 x 1ml 3 x 1ml 4 x 1ml
    Anti-FITC antibody HRP conjugate 1 x 100µl 2 x 100µl 3 x 100µl 4 x 100µl
    Bovine Serum Albumin 1 x 1g 2 x 1g 3 x 1g 4 x 1g
    Dry Skimmed milk 1 x 1g 2 x 1g 3 x 1g 4 x 1g
    Granzyme B Biotinylated detection antibody 1 vial 2 vials 3 vials 4 vials
    Granzyme B Capture Antibody 1 x 500µl 2 x 500µl 3 x 500µl 4 x 500µl
    Human IFN Gamma Capture antibody 1 x 500µl 2 x 500µl 3 x 500µl 4 x 500µl
    IFNγ FITC conjugated detection antibody 1 vial 2 vials 3 vials 4 vials
    Ready-to-use BCIP/NBT substrate buffer 1 x 50ml 2 x 50ml 3 x 50ml 4 x 50ml
    Streptavidin - Alkaline Phosphatase conjugated 1 x 50µl 2 x 50µl 3 x 50µl 4 x 50µl

  • 研究分野

  • 関連性

    Interferon 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. Granzyme B is necessary for target cell lysis in cell-mediated immune responses. It cleaves after Asp. Seems to be linked to an activation cascade of caspases (aspartate-specific cysteine proteases) responsible for apoptosis execution. Cleaves caspase-3, -7, -9 and 10 to give rise to active enzymes mediating apoptosis.

  • 別名

    • CSPB
    • Cytotoxic T lymphocyte proteinase 2
    • Granzyme 2
    • Granzyme B
    • GZMB
    • HLP
    • IFI
    • IFN gamma
    • IFNG
    • Immune interferon
    see all

  • 参照データベース

    アプリケーション

    The Abpromise guarantee

    Abpromise保証は、 次のテスト済みアプリケーションにおけるab48463の使用に適用されます

    アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

    アプリケーション Abreviews 特記事項
    ELISpot
    Use at an assay dependent dilution.
    特記事項
    ELISpot
    Use at an assay dependent dilution.

    参考文献 (0)

    ab48463 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab48463 は論文での使用が確認できていません。

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