Flow Cytometric Quantitation of Cellular Proteins
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Quantitation of specific proteins in complex mixtures is simplified by the use of antibodies directed against the protein of interest. If the specific protein is differentially expressed within a population of cells, quantitation of the protein in cell lysates by immunoblotting will provide an average quantity of the protein per cell. As a result, when comparing lysates of different cell populations, large changes in the amount of a protein in a small percentage of cells cannot be distinguished from small changes in the amount of the protein in a large percentage of cells. Flow cytometric analysis solves this problem by providing a means to measure the amount of a protein within each individual cell in a population. One restriction is that a specific fluorescently labeled probe, usually an antibody, is required for detection of the protein. Cells that are reacted with the antibody are then passed through a flow cytometer where a single cell suspension is focused into a stream that intersects a laser beam. As each cell passes through the beam, the fluorescent probes in each cell are excited and photomultiplier tubes register the degree of fluorescence (1).
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- 1.Shapiro, H. M. (1995) Practical Flow Cytometry. Wiley-Liss, New York.Google Scholar