Protein Quantitation Using Flow Cytometry
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It is often desirable to know how much of a particular protein is produced by a particular cell. For example, is this protein expressed at low levels throughout a population of cells, or is this protein expressed at very high levels in some cells and not in others? Does the expression of the protein change in a particular cell-cycle compartment? Technical protocols that require cell lysis, such as immunoblotting (1), allow the mean expression level of a given protein to be compared between populations of cells. It is even possible to use the immunoblotting protocol to determine the amount of a given protein expressed by a population of cells, if the protein has been purified for use as a standard. However, cell lysis obliterates the potential for single-cell quantitation of proteins.
KeywordsPropidium Iodide Repeat Centrifugation Flow Cytometer System Target Protein Molecule Minimize Cell Loss
- 2.Shapiro, H. M. (1995) Practical Flow Cytometry. Wiley-Liss, New York.Google Scholar