Rat IFN-gamma ELISpot Kit

Rat IFN-gamma ELISpot Kit

Assay Genie ELISpot is a highly specific immunoassay for the analysis of IFN-γ 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 IFN-γ 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 IFN-γ (qualitative analysis) and single cells that produce IFN-γ (quantitative analysis). Cell secreted IFN-γ is captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured IFN-γ is revealed by tracer antibodies and appropriate conjugates.

Rat IFN-gamma ELISpot Kit

The Rat IFN-gamma ELISPOT Kit is a powerful tool for detecting and quantifying Interferon-gamma levels in rat samples. This kit offers high sensitivity and specificity, providing accurate and reliable results for a variety of research applications.Interferon-gamma is a key cytokine involved in the immune response, playing a vital role in both innate and adaptive immunity. It is essential for host defense against infections, tumor surveillance, and autoimmune responses. By measuring IFN-gamma levels, researchers can better understand immune responses and identify potential therapeutic targets for a range of diseases and conditions.

With the Rat IFN-gamma ELISPOT Kit, researchers can easily analyze IFN-gamma production at the single-cell level, allowing for precise measurement of immune responses in experimental settings. This kit is an invaluable tool for studying immune function, vaccine development, and immunotherapy research in rat models.

A capture antibody highly specific for IFN-γ 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 IFN-γ produced. Cells are then removed by washing prior to the addition of Biotinylated detection antibodies which bind to the previously captured IFN-γ. 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.