Abstract
In general, most serodiagnostic assays offer an indirect measure of antibody reactivity with antigenic components, and the specific antigen for recognition is not absolutely clear unless a purified component antigen has been used. Certainly in the EIA format, it is generally assumed that a positive assay, which is indicated usually by a colour change, is directly reflective of an antigen-antibody interaction. Immunoblotting has the potential to enhance serodiagnosis by allowing the observer to determine the specific antigen which is being recognized by the antibody response. Antigens are initially separated by an electrophoresis technique such as reducing or non-reducing sodium dodecyl sulphate Polyacrylamide gel electrophoresis (SDSPAGE). The resulting polypeptide profile is then transferred to a solid support (e.g. nitrocellulose) where the enzyme-linked antibody reaction can occur and where it can be visualized. Reactivity with single or multiple antigens is qualitatively determined although IgM detection, for example, could be determined on the basis of total absence (negative) or the presence of any quantity (positive). Although the resolution of antigen is commonly in the form of a one dimensional ladder, two dimensional resolution is also possible. The antigens which are resolved in this manner are believed to be mainly polypeptides and indeed, CF glycolipid antigens are not capable of absorbing out antibody to the latter.157
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© 1999 Springer Science+Business Media New York
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Cimolai, N. (1999). Immunoblotting. In: Serodiagnosis of the Infectious Diseases. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-5249-9_12
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DOI: https://doi.org/10.1007/978-1-4615-5249-9_12
Publisher Name: Springer, Boston, MA
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