What is the principle behind Elisa spot?
The Elisa spot technique was originally described by Sedgwick & Holt, Journal of Immunological Methods, 57, 301, 1983. The basic principle is as follows: • Coat solid phase with antigen • Block free sites using serum • Add antibody producing cells (plasma cells) from animal, e.g. mouse sensitized with the coated antigen • Incubate • Wash away cells • Add anti-mouse antibody conjugated with alkaline phosphatase • Incubate • Wash • Add substrate • Incubate • Read number of spots (converted substrate) Elisa spot can also be used to assay products secreted from cells placed in contact with antibody coated on the solid phase.
