Immunoblotting techniques.
Immunoblotting techniques use antibodies (or other specific ligands in related techniques) to identify target proteins among a number of unrelated protein species. They involve identification of protein target via antigen-antibody (or protein-ligand) specific reactions. Proteins are typically separated by electrophoresis and transferred onto membranes (usually nitrocellulose). The membrane is overlaid with a primary antibody for a specific target and then with a secondary antibody labeled, for example, with enzymes or with radioisotopes. When the ligand is not an antibody, the reaction can be visualized using a ligand that is directly labeled. Dot blot is a simplified procedure in which protein samples are not separated by electrophoresis but are spotted directly onto membrane. Immunoblotting is now widely used in conjunction with two-dimensional polyacrylamide gel electrophoresis, not only for traditional goals, such as the immunoaffinity identification of proteins and analysis of immune responses but also as a genome-proteome interface technique.
Magi
Barbara
B
Department of Molecular Biology, University of Siena, Siena, Italy.
Liberatori
Sabrina
S
D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D015153:Blotting, Western / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D008567:Membranes, Artificial; D020543:Proteome
D000906:Antibodies; D008567:Membranes, Artificial; D020543:Proteome