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Your Position: Home > Kits > IL-4 > RAS-SP010

Mouse IL-4 ELISPOT Kit

For research use only.

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    Materials Provided
    IDComponentsSize
    RSP010-C01Pre-coated Anti-IL-4 Antibody Microplate1 plate
    RSP010-C02Positive Stimulus150 μg
    RSP010-C03Biotin-Anti-IL-4 Antibody50 μL
    RSP010-C04Streptavidin-HRP50 μL
    RSP010-C05Washing Buffer (10×)50 mL
    RSP010-C06Dilution Buffer (1×)50 mL
    RSP010-C07AEC Dilution 25 mL
    RSP010-C08AEC Solution A (20×)0.8 mL
    RSP010-C09AEC Solution B (20×)0.8 mL
    RSP010-C10AEC Solution C (20×)0.8 mL
  • Background
    IL-4 is known to be the quintessential regulatory cytokine, playing a role in a vast number of immune and non-immune functions. This cytokine is commonly secreted by type 2 helper T (TH2) cells and follicular helper T (TFH) cells after antigenic sensitization. The Mouse IL-4 ELISpot assay is designed for the detection of mouse IL-4 secreting cells at the single cell level and can be used to quantitate the frequency of mouse IL-4 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 mouse IL-4 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-4 Antibody Microplate, Positive Stimulus, Biotin-Anti-IL-4 Antibody, Streptavidin-HRP, AEC and buffers.

    Your experiment will include 6 simple steps:

    a) A monoclonal antibody specific for mouse IL-4 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-4.

    c) After washing away any cells and unbound substances, a biotinylated antibody specific for mouse IL-4 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-4 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-4 TYPICAL DATA

2.5×10⁵ cells/well Mouse Spleen Cells were stimulated with 18 μg/mL Positive Stimulus and cultured at 37℃ and 5%CO2 for 20h, 228 spots were produced. The following example data is for reference only.

  • Clinical and Translational Updates

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Price(USD) : $298.00

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