IL-12p70 ELISpot Assay Kit


This IL-12p70 ELISpot Assay Kit is designed to determine the frequency of IL-12p70 producing cells under a given stimulation and the comparison of such frequency against a specific treatment or pathological state. The Eagle Biosciences IL-12p70 ELISpot Assay Kit is for research use only and not to be used for diagnostic procedures.

IL-12p70 ELISpot Assay Kit

The IL-12p70 ELISpot Assay Kit is for research use only.

Incubation Time: 18hrs – 23hrs
Sample Size: 100 µl
Alternative Names: Interleukin-12p70

Assay Principle for IL-12p70 ELISpot

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 level in conditions closely comparable to the in-vivo environment with minimal 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 to IL-12p70

IL-12p40 ELISA Assay Kit
Human IL-12p70 ELISA Assay
iLite IL-12 Assay Ready Cells

Additional Information

Assay Background

IL-12 is a potent regulator of cell mediated immune response produced by activated monocytes / macrophages cells, B lymphocytes and connective tissue type mast cells. The biologically active form of IL-12 is a 70 kDa heterodimeric glycoprotein consisting of disulfide-linked 35 kDa (p35) light chain and 40 kDa (p40) heavy chain subunits. The two subunits are genetically unrelated. The p70 form is the only biologically active form of IL-12. The p35 subunit has homology to IL-6, while p40 has homology with IL-23. IL-12 has been found to bind to IL-12R. IL-12R has been reported to be present on IL-2 activated CD4+, CD8+ and CD56+ cells. IL-12 exerts a variety of biological effects on human T and NK cells. IL-12 induces an IFNγ production and other cytokines from peripheral blood T and NK cells. Its role is directing development and proliferation of Th1 cells. Thus IL-12 is linked with autoimmunity, high level have also been reported for chronic inflammatory reactions, bacterial and viral infection.

ELISpot Assay Kit Procedure


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