Introduction to EliSpot (Enzyme Linked Immunospot ) Technology
Elispot technique (Enzyme Linked Immunospot Assay, ELISPOT) is one of the cellular immunology of the most sensitive detection method, and cytokines (CK) secreting cells were detected on the antibody-secreting cells (ASC) the single cell level. Because it is a single cell detection level, the ELISPOT and ELISA than the limiting dilution method or the like has a higher sensitivity, from 200,000 to 300,000 cells detected in one of the secreted protein in the cell. Moreover, the method can perform functional detection on living cells after antigen stimulation, has high specificity, high visual credibility, and is easy to operate, and has been widely used in secretion of CK cell detection or ASC measurement, at home and abroad. The immunology community has gained wide application.
Principle of ELISPOT Principle :
Upon stimulation of the cells, cytokines are produced, which are captured by specific monoclonal antibodies on the PVDF membrane of the ELISPOT plate . Upon removal of the cells, the captured cytokine binds to the biotinylated secondary antibody and thereafter binds to alkaline phosphatase or horseradish peroxidase-labeled avidin. After incubation of the substrate, purple or reddish-brown spots on the PVDF wells indicate that the cells produced cytokines, and the spots were automatically imaged, counted, and analyzed by an AID ELISPOT analyzer (enzyme-linked spot analysis system).
ELISPOT Protocol method steps :
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Why choose ELISPOT assay?
•    High sensitivity: one millionth of positive cells , 200x vs ELISA, ~RT-PCR
•    Live cell function test: The detected cytokine is not already present, but is secreted by the stimulated stimulus during the test.
•    Intuitive and highly reliable: one positive cell has a spot, the number of spots represents the number of cells that secrete cytokines, and the size of the spot represents the level of cytokine secretion by the cell.
•    Cells that have been cryopreserved do not lose their immune function: this is very important for clinical trials, and pre- and post-treatment samples can be tested side-by-side and the results can be repeated;
•    High-throughput testing: Researchers can test hundreds of samples per day; only a small amount of cells, such as 45 ml of blood, is sufficient to test the response of 600 different antigen / peptide species.
In your immunology research, ELISPOT is an effective research tool for detecting cytokines secreted by antigen-specific T cells or antibodies secreted by B cells for T and B cell function studies. For more information on ELISPOT experiments and techniques, visit
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