The most common method for analyzing protein expression levels is Western blotting with detection of a single specific protein target, using horseradish peroxidase– or alkaline phosphatase–conjugated antibody probes combined with colorimetric or chemilumines-cent detection. However, multiplex analyses using these techniques are time-consuming and imprecise.
Western blotting is a widely used and sensitive technique for the detection and characterization of proteins. This technique takes advantage of antibody recognition specificity toward an antigenic sequence in a target protein.
Western blot transfer from an SDS-PAGE gel is a method that can be used to detect individual proteins in a given complex sample. The denatured proteins are resolved by electrophoresis then transferred out of the PAGE gel and onto a microporous membrane, where they are located by stains or by “probing” with specific detection reagents, such as antibodies to the analyte of interest.
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