製品の概要

  • 製品名
    Rat Interferon gamma ELISPOT Kit
    Interferon gamma キット 製品一覧
  • 検出方法
    Colorimetric
  • サンプルの種類
    Cell culture supernatant, Cell culture extracts
  • アッセイタイプ
    Sandwich (qualitative)
  • ステップ
    Multiple steps standard assay
  • 種交差性
    交差種: Rat
  • 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, cancerology, infectious diseases, autoimmune diseases and transplantation.

    Abcam 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 rat Interferon gamma.

  • アプリケーション
    適用あり: ELISpotmore details
  • 試験プラットフォーム
    Microplate

製品の特性

  • 保存方法
    Store at +4°C. Please refer to protocols.
  • 内容 1 x 96 tests 5 x 96 tests
    Bovine Serum Albumin 1 x 2g 1 x 2g
    IFNγ Biotinylated Detection Antibody 1 x 100µl 1 vial
    Rat IFNγ Pre-coated 96-well PDVF-bottomed plates 2 units 5 units
    Ready-to-use BCIP/NBT substrate buffer 1 x 11ml 2 x 25ml
    Streptavidin - Alkaline Phosphatase conjugated 1 x 10µl 1 x 50µl
  • 研究分野
  • 関連性
    Interferon gamma is a dimerized soluble cytokine that is the only member of the type II class of interferons. In contrast to interferon alpha and interferon beta which can be expressed by all cells, Interferon gamma is secreted by T lymphocytes and NK cells only. Also known as immune interferon, Interferon gamma is the only Type II interferon. It is serologically distinct from Type I interferons and it is acid-labile, while the type I variants are acid-stable. Interferon gamma has antiviral, immunoregulatory, and anti-tumour properties. It alters transcription in up to 30 genes producing a variety of physiological and cellular responses. Activation by Interferon gamma is achieved by its interaction with a heterodimeric receptor consisting of IFNGR1 & IFNGR2 (interferon gamma receptors). Interferon gamma binding to the receptor activates the JAK-STAT pathway. In addition, Interferon gamma activates APCs and promotes Th1 differentiation by upregulating the transcription factor T-bet. Interferon gamma is the hallmark cytokine of Th1 cells (Th2 cells produce IL-4). NK cells and CD8+ cytotoxic T cells also produce Interferon gamma. Interferon gamma suppresses osteoclast formation by rapidly degrading the RANK adaptor protein TRAF6 in the RANK-RANKL signaling pathway, which otherwise stimulates the production of NF?B.
  • 細胞内局在
    Secreted
  • 別名
    • IFG
    • IFI
    • IFN gamma
    • IFNG
    • Immune interferon
    see all
  • 参照データベース

アプリケーション

Our Abpromise guarantee covers the use of ab64032 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

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

参考文献

ab64032 has not yet been referenced specifically in any publications.

レビューと Q&A

Delivery plus Positive Control Issues

Below average Good 4/5 (Ease of Use)
Abreviews
I ordered this product in July when Abcam's site said a 1-2 week lead time on delivery. I received the product roughly 4 weeks later after several emails apologizing for the product's delay.

I unsuccessfully used the first 3 plates (following each step in the instruction book carefully) on freshly isolated rat PBMCs and splenocytes, without any spots on my positive controls of varying PMA + ionomycin concentrations. Abcam recommends PMA + ionomycin at 1 and 500 ng/mL in the instruction booklet, but PHA at 10 ug/ml here: http://www.abcam.com/protocols/elispot-protocol. I decided to try PHA at this concentration and my spots appeared beautifully, with nothing on the PMA+ionomycin wells.

Between the delay and the instruction booklet setback, I have to give this product 2 stars.
Username

Benjamin Maimon

Verified customer

投稿 Oct 25 2017

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

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