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mskyeg Group Title

Antibodies can be used to find, bind, and tag specific molecules of interest. Antibodies are Y-shaped molecules, with the tips of the Y containing unique amino acid sequences that bind the antigen. These variable portions of antibody molecules convey the high specificity for a target molecule. The large tail or base of the Y is much less variable. In fact, all antibodies within a species have tail regions that are very similar in sequence and shape. By inserting compounds that fluoresce, meaning they produce radiation or produce a color change in the tail region of these molecules, researchers

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    can use antibodies as marker molecules. Researchers often use two different antibodies: a primary antibody for targeting the molecule of interest and a secondary antibody with active sites to bind the primary antibody’s tail and act as a marker. Western blots are used to detect the presence of a known protein in a given sample. The proteins are first separated by molecular weight using gel electrophoresis. Next, the proteins are transferred from the gel to a nitrocellose membrane in a process called blotting. The nitrocellulose membrane is then incubated with a primary antibody that combines with the protein of interest. Then an enzyme coupled with a secondary antibody is used to produce a detectable color change when the protein of interest is present. The intensity of the color change indicates the quantity of the protein. Immunohistochemistry uses antibodies to detect the presence of specific molecules, usually proteins, within tissues and cells. Thin sections of a biological sample are fixed to a glass slide, incubated with primary and secondary antibodies, and examined microscopically. Secondary antibodies used in immunohistochemistry are frequently fluorescent, in which case a fluorescent microscope is used to read the results. Briefly describe what you can learn about a target protein by using these two techniques.

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