Human Fas Ligand ELISpot Kit

Human Fas Ligand ELISpot Kit

Assay Genie ELISpot is a highly specific immunoassay for the analysis of Fas Ligand 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 Fas Ligand producing cells under a given stimulation and the comparison of such frequency against a specific treatment or pathological state. Utilising sandwich immuno-enzyme technology, Assay Genie ELISpot assays can detect both secreted Fas Ligand (qualitative analysis) and single cells that produce Fas Ligand (quantitative analysis). Cell secreted Fas Ligand is captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured Fas Ligand is revealed by tracer antibodies and appropriate conjugates.

Human Fas Ligand ELISpot Kit

The Human FAS Ligand ELISPOT Kit is a cutting-edge tool designed for the precise quantification of FAS Ligand levels in human samples, including serum, plasma, and cell culture supernatants. This kit boasts exceptional sensitivity and specificity, ensuring accurate and consistent results that are essential for a wide range of research applications.FAS Ligand, also known as CD178, is a critical protein involved in programmed cell death and immune regulation. Its dysregulation has been linked to various diseases including autoimmune disorders, cancer, and inflammatory conditions.

Therefore, the accurate measurement of FAS Ligand levels is crucial for understanding disease pathogenesis and identifying potential therapeutic targets.With the Human FAS Ligand ELISPOT Kit, researchers can confidently study the role of FAS Ligand in health and disease, paving the way for the development of novel diagnostic tools and targeted therapies. Experience the precision and reliability of this advanced kit for your research needs.

A capture antibody highly specific for Fas Ligand is coated to the wells of a PVDF bottomed 96 well microtitre 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 Fas Ligand produced. Cells are then removed by washing prior to the addition of Biotinylated detection antibodies which bind to the previously captured Fas Ligand. 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.

Figure: Schematic representation of the principle of ELISpot. 1) Treat with ethanol, coat with capture antibody. 2) Incubate cells in the presence of stimuli, cytokines, are released, which bind to capture antibodies. 3) Wash plate. Incubate with biotinylated detection antibody. 4) Wash plate. Incubate with streptavidin-alkaline phosphatase conjugated. 5)Incubate with BCIP/NBT. Precipitated product forms, results in blue/purple spots.