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Your Position: > Kits > IL-2 > RAS-SP006

Mouse IL-2 ELISPOT Kit

For research use only.
    Materials Provided
    IDComponentsSize
    RSP006-C01Pre-coated Anti-IL-2 Antibody Microplate1 plate
    RSP006-C02Positive Stimulus60 μg
    RSP006-C03Biotin-Anti-IL-2 Antibody50 μL
    RSP006-C04Streptavidin-HRP50 μL
    RSP006-C05Washing Buffer (10×)50 mL
    RSP006-C06Dilution Buffer (1×)50 mL
    RSP006-C07AEC Dilution 25 mL
    RSP006-C08AEC Solution A (20×)0.8 mL
    RSP006-C09AEC Solution B (20×)0.8 mL
    RSP006-C10AEC Solution C (20×)0.8 mL
  • Background
    Interleukin-2 (IL-2) is a pleiotropic cytokine that drives T-cell growth, augments NK cytolytic activity, induces the differentiation of regulatory T cells, and mediates activation-induced cell death. IL-2 does not specify the type of Th differentiation that occurs; instead, IL-2 modulates expression of receptors for other cytokines and transcription factors, thereby either promoting or inhibiting cytokine cascades that correlate with each Th differentiation state. The Mouse IL-2 ELISpot assay is designed for the detection of mouse IL-2 secreting cells at the single cell level and can be used to quantitate the frequency of mouse IL-2 secreting cells. ELISpot assays are highly reproducible and sensitive, and can be used to measure responses with frequencies well below 1 in 100,000. ELISpot assays do not require prior in vitro expansion of T cells and they are suitable for high-throughput analysis using only small volumes of primary cells. As such, ELISpot assays are useful tools for research in vaccine development, cytokine secretion and the monitoring of various clinical trials.
  • Application

    This kit is used to detect IL-2 at the cellular level.

    It is for research use only.

  • Reconstitution
    Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
  • Storage
    1. Unopened kit should be stored at 2℃-8℃ upon receiving.

    2. Find the expiration date on the outside packaging and do not use reagents past their expiration date.

    3. Please use it up once you opened it.

  • Assay Principles
    ELISpot assays employ the quantitative sandwich enzyme-linked immunosorbent assay (ELISA) technique. The Kit consists of Pre-coated Anti-IL-2 Antibody Microplate, Positive Stimulus, Biotin-Anti-IL-2 Antibody, Streptavidin-HRP, AEC and buffers.

    Your experiment will include 6 simple steps:

    a) A monoclonal antibody specific for mouse IL-2 has been pre-coated onto a polyvinylidene difluoride (PVDF)-backed microplate.

    b) Appropriately stimulated cells are pipetted into the wells and the microplate is placed into a humidified 37 °C CO2 incubator for a specified period of time.During this incubation period, the immobilized antibody in the immediate vicinity of the secreting cells binds secreted IL-2.

    c) After washing away any cells and unbound substances, a biotinylated antibody specific for mouse IL-2 is added to the wells. Following a wash to remove any unbound biotinylated antibody, HRP conjugated to streptavidin is added.

    d) Wash the plate and add the Streptavidin-HRP diluted by Dilution Buffer to the plate.

    e) Wash the plate and add AEC. A reddish-brown colored precipitate forms and appears as spots at the sites of cytokine localization, with each individual spot representing an individual IL-2 secreting cell.

    f) The spots can be counted with an automated ELISpot reader system or manually using a stereomicroscope.

Typical Data Please refer to DS document for the assay protocol.
 IL-2 TYPICAL DATA

1×10⁵ cells/well Mouse Spleen Cells were stimulated with 6.0 μg/mL Positive Stimulus and cultured at 37℃ and 5%CO2 for 16 h, 265 spots were produced. The following example data is for reference only.

  • Clinical and Translational Updates

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