Overview

  • Product nameHuman IL17A/F ELISPOT Kit
    See all IL17A/F kits
  • Detection methodColorimetric
  • Tests
    15 x 96 well plate
  • Sample type
    Cell culture supernatant, Cell culture extracts
  • Assay typeSandwich (qualitative)
  • Assay durationMultiple steps standard assay
  • Product overview

    ab105247 is a highly specific immunoassay for the analysis of cytokine and other soluble molecule production and secretion from T-cells at a single cell level in conditions closely comparable to the in-vivo environment with minimal cell manipulation. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation and the comparison of such frequency against a specific treatment or pathological state. The ELISpot assay constitutes an ideal tool in the investigation of Th1 / Th2 responses, vaccine development, viral infection monitoring and treatment, oncology, infectious disease, autoimmune diseases and transplantation.

     

    Utilising sandwich immune-enzyme technology, ELISpot assays can detect both secreted cytokines and single cells that simultaneously produce multiple cytokines. 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.

    A capture antibody highly specific for the molecule of interest is coated to the wells of a PVDF bottomed 96 well microtiter plate either during kit manufacture or in the laboratory. The plate is then blocked to minimise any non-antibody dependent unspecific binding and washed. Cell suspension and stimulant are added and the plate incubated allowing the specific antibodies to bind any molecules produced. Cells are then removed by washing prior to the addition of Biotinylated detection antibodies which bind to the previously captured molecule. Enzyme conjugated streptavidin is then added binding to the detection antibodies. Following incubation and washing substrate is then applied to the wells resulting in coloured spots which can be quantified using appropriate analysis software or manually using a microscope. See Figure 1 for visual scheme.

     

    ab105247 provides enough reagents for 15 plates. Plates are provided and are not sterile.
     

    The assay recognizes natural human IL17A + IL17F. After testing, no cross reactivity was observed for IL17A, IL17B, IL17D, IL17E, IFN-g, Gal-1 and IL33. This antibody pair shows cross reactivity with human IL17F.

  • Notes
    • Store kit reagents between 4°C.
    • uncoated plates should be stored at RT.

     

    • Immediately after use remaining reagents should be returned to cold storage (4°C).
    • All reagents should be warmed to room temperature before use.
    • BCIP/NBT buffer is potentially carcinogenic and should be disposed of appropriately, caution should be taken when handling this reagent, always wear gloves
    • Capture Antibody is supplied sterile: once opened keep the vial sterile or aliquot and store at -20oC. For optimal performance prepare the Capture Antibody dilution immediately before use.
    • If not used within a short period of time, reconstituted Detection Antibody should be aliquoted and stored at -20°C. In these conditions the reagent is stable for at least one year.

     

    • For optimal performance prepare the Streptavidin-AP dilution immediately prior to use. Do not keep this solution for further experiments.

     

     

     

     

     

  • Tested applicationsSuitable for: ELISpotmore details
  • PlatformMicroplate

Properties

    Applications

    Our Abpromise guarantee covers the use of ab105247 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.

    Human IL17A/F ELISPOT Kit images

    Protocols

    References for Human IL17A/F ELISPOT Kit (ab105247)

    ab105247 has not yet been referenced specifically in any publications.

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