Visualizations of the different secretions of cells that are individually activated or responding is possible using ELISPOT assays, provided by Abcam. Additionally, the data provided can be qualitative, to see if particular proteins are present, or quantitative, to determine the level of protein response.
The target cells are grown on plates that are coated in antibodies. Once the cells are stimulated, these antibodies respond to the cell secretions. Using a sandwich ELISA method, the distribution of secreted analytes can be determined easily as a visualized reporter substance is produced upon antibody:antigen capture.
Every spot produced over the course of the assay corresponds to a reactive cell. This provides rich data that is both quantitative, as the volume of cells responding can be measured, and qualitative, as individual immune proteins can be identified.
FLUROSPOT assays use a sandwich ELISA method but with a fluorescent, rather than chromogenic, detection method.
FLUROSPOT assays can be used to identify which cells are secreting proteins and are especially useful for analyzing the secretion of soluble molecules such as cytokines from T-cells.
Find Out More About Dual Staining
Dual staining versions of ELISPOT and FLUROSPOT allow the secretion of two different proteins, and the frequency of their secretion, from single cells to be measured at the same time. This prevents researchers from having to run multiple assays and can allow researchers to determine if cytokines are co-expressed.
The Different Detection Modes of Dual Flurospot and Dual Elispot
Dual FLUROSPOT uses anti-FITC antibody green fluorescence and streptavidin-phycoerythrin conjugates.
Dual ELISPOT uses an AEC substrate buffer (red spots) and BCIP/NBT (blue spot), both of which can be used immediately.
Abcam is a global life sciences company providing highly validated antibodies and other binders and assays to the research and clinical communities to help advance the understanding of biology and causes of disease.
Abcam’s mission is to serve life scientists to help them achieve their mission faster by listening to their needs, continuously innovating and improving and by giving them the tools, data and experience they want. Abcam’s ambition is to become the most influential life science company for researchers worldwide.
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