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Flow Cytometric Quantitation of Cellular Proteins

  • Thomas D. Friedrich
  • F. Andrew Ray
  • Judith A. Laffin
  • John M. Lehman
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

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).

Keywords

Propidium Iodide Light Scatter Wash Solution Resuspend Pellet African Green Monkey Kidney Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Thomas D. Friedrich
    • 1
  • F. Andrew Ray
    • 2
  • Judith A. Laffin
    • 3
  • John M. Lehman
    • 4
  1. 1.Center for Immunology and Microbial DiseaseAlbany Medical College
  2. 2.Department of BiologyHartwick CollegeOneonta
  3. 3.Department of Microbiology, Immunology, and Molecular GeneticsThe Albany Medical CollegeAlbany
  4. 4.Center for Immunology and Microbial DiseaseAlbany Medical College

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