Detection of Polypeptides on Immunoblots Using Enzyme- Conjugated or Radiolabeled Secondary Ligands
2003; Humana Press; Linguagem: Inglês
10.1385/1-59259-169-8
Autores Tópico(s)Glycosylation and Glycoproteins Research
ResumoImmunoblotting provides a simple and effective method for identifying specific antigens in a complex mixture of proteins. Initially, the constituent polypeptides are separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDSPAGE), or a similar technique, and are then transferred either electrophoretically or by diffusion onto a nitrocellulose filter. Once immobilized on a sheet of nitrocellulose, specific polypeptides can be identified using antibodies that bind to antigens retained on the filter and subsequent visualization of the resulting antibody-antigen complex. This chapter describes conditions suitable for binding antibodies to immobilized proteins and methods for locating these antibody-antigen complexes using appropriately labeled ligands. These methods are based on those of Blake et al. (), Burnette () and Towbin et al. ().
Referência(s)