Mouse IFN gamma ELISpot Assay Kit

$350.00$1,550.00

This Mouse IFN gamma ELISpot Assay Kit is designed to determine the frequency of IFN gamma producing cells under a given stimulation and the comparison of such frequency against a specific treatment or pathological state. The Eagle Biosciences Mouse IFN gamma ELISpot Assay Kit is for research use only and not to be used for diagnostic procedures.

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Mouse IFN gamma ELISpot Assay Kit

The Mouse IFN gamma ELISpot Assay Kit is for research use only.

Incubation Time: 18hrs – 23hrs
Sample Size: 100 µl


Assay Principle

The ELISpot 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, cancerology, infectious disease, autoimmune diseases and transplantation. Utilizing sandwich immuno-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 analyte 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 minimize 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 analytes produced. Cells are then removed by washing prior to the addition of Biotinylated detection antibodies which bind to the previously captured analyte. Enzyme conjugated streptavidin is then added binding to the detection antibodies. Following incubation and washing substrate is then applied to the wells resulting in colored spots which can be quantified using appropriate analysis software or manually using a microscope.


Related Products

High Sensitive IFN Gamma ELISA Assay Kit
IFN gamma ELISpot Assay Kit

Additional Information

Assay Background


Different populations of T-cells secrete differing patterns of cytokines that ultimately lead to different immune responses. IFNγ production is a key function of Th1, CD8+ CTLs and also NK cells. IFNγ is a cytokine critical for cell mediated immunity against viral and intracellular bacterial infections and is involved in the inflammatory response following secretion via macrophage activation and stimulation of antibody secretion. IFNγ is the hallmark effector cytokine of Th1 and therefore is an excellent marker for identifying a host response to intracellular pathogens.
IFNγ is produced during infection by T cells of the cytotoxic/suppressor phenotype (CD8) and by a subtype of helper T cells, the Th1 cells. Th1 cells secrete IL-2, IL-3, TNFα and IFNγ, whereas Th2 cells mainly produce IL-3, IL-4, IL-5, and IL-10 but little or no IFN-γ. IFNγ preferentially inhibits the proliferation of Th2 but not Th1 cells, indicating that the presence of IFNγ during an immune response will result in the preferential proliferation of Th1 cells. In addition, IFNγ has several properties related to immunoregulation. IFNγ is a potent activator of mononuclear phagocytes, and activates macrophages to kill tumor cells by releasing reactive oxygen intermediates and TNFα. IFNγ induces or augments the expression of MHC antigens on macrophages, T and B cells and some tumor cell lines. On T and B cells IFNγ promotes differentiation. It enhances proliferation of activated B cells and can act synergistically with IL-2 to increase immunoglobulin light-chain synthesis. The role of IFNγ as a disease marker has been demonstrated for a number of different pathological situations including, viral infection, Autoimmune disease, transplant rejection, Diabetes and allergy.

ELISpot Assay Kit Procedure

Documents

Product Manual