IL-1 beta ELISpot Assay Kit


This IL-1 beta ELISpot Assay Kit is designed to determine the frequency of IL-1 beta producing cells under a given stimulation and the comparison of such frequency against a specific treatment or pathological state. The Eagle Biosciences IL-1 beta ELISpot Assay Kit is for research use only and not to be used for diagnostic procedures.

IL-1 beta ELISpot Assay Kit

The IL-1 beta 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

IL-1 Beta ELISA Assay Kit
Mouse IL-1 Best ELISA Assay Kit

Additional Information

Assay Background

Interleukin-1 Beta (IL-1β) is a member of the interleukin-1 family. This family contains three structurally related polypeptides. The first two are IL-1α and IL-1β, each of which has a broad spectrum of both beneficial and harmful biologic actions, and the third is IL-1-receptor antagonist, which inhibits the activities of interleukin-1. IL-1α and β present approximately 25% homology at the amino acid level, but the difference is in their tri dimensional structure. Two distinct receptor types have been isolated, that bind both forms. IL-1β is synthesized as a large precursor, with a molecular weight of 31 kDa. The molecular weight of the mature form is 17.5 kDa. Unlike IL-1α, the IL-1β precursor shows little or no biological activity in comparison to the mature form. IL-1 is primarily an inflammatory cytokine. It belongs to a group of cytokines with overlapping biologic properties (TNFα and IL-6). IL-1, TNF and IL-6 share the ability to stimulate T and B lymphocytes, increase cell proliferation, and initiate or suppress gene expression for several proteins exerting their effects by binding to specific receptors. IL-1 (α and β) have similar biological properties, among them, the ability to induce fever, sleep, anorexia and hypotension. IL-1 stimulates the release of pituitary hormones, increases the synthesis of collagenases, resulting in the destruction of cartilage, and stimulates the production of prostaglandins, leading to decrease in the pain threshold. In addition IL-1 has some host defence properties. However, whereas IL-1β is a secreted cytokine, IL-1α is predominantly a cell-associated cytokine. IL-1 has also been implicated in the destruction of beta cells of the islets of Langerhans, the growth of myelogenous leukaemia cells, and the development of atherosclerotic plaques. It is described in several diseases: sepsis syndrome, rheumatoid arthritis, inflammatory Bowel disease, acute and chronic myelogenous leukaemia, insulin dependent diabetes mellitus and arthrosclerosis.

ELISpot Assay Kit Procedure


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