Structural Biochemistry/Enzyme-Linked ImmunoSpot Assay (ELISpot)

Overview
A modified version of the ELISA immunoassay, the ELISpot or enzyme-linked immunosorbent spot, allows for the detection of cells that secrete cytokines or antibodies. Unlike ELISA, ELISpot is able to detect a single cell that secretes a protein of interest making it one of the most sensitive cellular assays.

A quick and simplified procedure is listed below: 1) Cytokine-specific antibodies are placed onto an ELISpot plate 2) Cells of interest are then added 3) Cells become activated and begin to produce cytokine which binds to the antibodies 4) The cells are then removed whilst detection antibodies that can bind with the enzyme are added 5) Finally a substrate forms a colored spot the secreting cell 6) The cells are then counted through a microscope