Final answer:
Western Blot analysis is used to detect specific proteins using antibodies and is semi-quantitative. It involves an enzymatic reaction that visualizes the proteins, but a colorimetric assay is used to determine protein concentration.
Step-by-step explanation:
Western Blot analysis is a widely used method in molecular biology for detecting specific proteins in a sample.
The technique involves separating proteins via polyacrylamide gel electrophoresis (SDS-PAGE), transferring them onto a nitrocellulose or PVDF membrane, and using antibody probes for detection.
The primary antibody binds to the protein of interest, and a secondary antibody with a molecular beacon, such as an enzyme or a fluorophore, binds to the primary antibody allowing visualization.
Although Western Blot can provide semi-quantitative data on protein expression levels, it does not directly determine the concentration of a protein.
Instead, protein concentrations in samples are often measured using a colorimetric assay to create a standard curve, as the color intensity correlates with protein amount.
The function of the enzyme in a Western Blot is to catalyze a chromogenic reaction, producing a color or emitting light to visualize the protein of interest on the membrane.