INTRODUCTION

The chapter is part of a group of general information chapters for immunological test methods (Immunological Test Methods—General Considerations 〈1102〉, Immunological Test Methods—Enyzme-Linked Immunosorbent Assay (ELISA) 〈1103〉, and Immunological Test Methods—Surface Plasmon Resonance 〈1105〉) that provides analysts with general information about principles, method development, method validation, and data evaluation for immunoblot analysis. Immunoblot analysis is defined as any method in which an antibody is used for detection of one or more analytes (e.g., proteins, polysaccharides) that has been transferred to a test membrane surface. Immunoblot methods are typically classified by whether electrophoretic separation occurs as a part of the immunoblot procedure. Electrophoretic separation is based on molecular weights and charge differences of a population of molecules. See Capillary Electrophoresis 〈1053〉, Biotechnology-Derived Articles—Isoelectric Focusing 〈1054〉, and Biotechnology-Derived Articles—Polyacrylamide Gel Electrophoresis 〈1056〉 for a detailed description of electrophoretic separation methods. An example of immunoblot analysis involving electrophoretic separation is the Western blot, which was first described in the scientific literature in the late 1970s. Another approach for immunoblot analysis is to perform molecule detection using an antibody without prior electrophoretic separation. Examples of this nonelectrophoretic type of approach are the slot or dot blot (slot/dot).