IL-4 ELISpot Assay Kit

$350.00$1,550.00

This IL-4 ELISpot Assay Kit is designed to determine the frequency of IL-4 producing cells under a given stimulation and the comparison of such frequency against a specific treatment or pathological state. The Eagle Biosciences IL-4 ELISpot Assay Kit is for research use only and not to be used for diagnostic procedures.

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IL-4 ELISpot Assay Kit

The IL-4 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

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Additional Information

Assay Background


IL-4 is a lymphokine that co-stimulates the proliferation of activated B- and T-cells, augments the cytotoxic activity of lymphocytes and monocytes and enhances the functional activity of myeloid cells. Produced by mast cells, T-cells and bone marrow stromal cells, IL-4 regulates the differentiation of naive CD4+ cells into helper Th2 cells characterized by their cytokine secretion-profile that includes secretion of IL-4, IL-5, IL-6, IL-10 and IL-13 which favour a humoral immune response. In addition, IL-4 can inhibit the proliferation of TNF, IL-1 and IL-6 by macrophages. IL-4 induces B-cell class switching to IgE and IgG1 isotypes, and up regulates MHC Class II production and CD23 expression. IL-4 is a 15kDa globular glycoprotein containing 129 amino acid residues. The non-glycosylated form of the protein is fully biologically active.

ELISpot Assay Kit Procedure

Documents

Product Manual