Membranes and membrane plates used in ELISPOT

Membrane-bottomed, 96-well plates constitute the format in which the overwhelming majority of enzyme-linked immunospot (ELISPOT) assays are performed. The membranes in these plates are made from either nitrocellulose or polyvinylidene fluoride. These membranes are well suited for ELISPOT because they have high antibody binding capacities and because their white color provides an excellent backdrop for ELISPOT enumeration. These two membranes and, ultimately, the 96-well plates used in ELISPOT assays were commercialized for filtration applications and later optimized for deoxyribonucleic acid hybridization and protein chemistry applications. In this chapter, an overview of the development and biotechnology applications of nitrocellulose and polyvinylidene fluoride membrane is provided and characteristics and attributes of each of the membranes that are relevant to ELISPOT are summarized.