Abstract
To measure the quantity of tyrosine phosphorylated proteins in cells, we have used a flow cytometry technique with fluorescein isothiocyanate-labeled anti-phosphotyrosine antibody (FITC-αPY mAb). The analysis was applied to the phosphotyrosine titration and showed an optimum amount of FITC-αPY mAb (30μg/1×10cells). The staining specificity of our assay was tested by the addition of exogenous competitors, showing a specific inhibition by phosphotyrosine but not by phosphoserine, phosphothreonine, or tyrosine. The assay was also able to elucidate the inhibitory effect on tyrosine phosphorylation of genistein, a protein tyrosine kinase inhibitor. These results imply that immunofluorescent quantification assay using a flow cytometer could be a useful technique to determine the intracellular level of tyrosine phosphorylated protein.
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Yoon, D.Y., Yoon, S.Y., Koh, W.S. et al. Immunofluorescent quantification of tyrosine phosphorylated proteins by flow cytometric analysis. Biotechnology Techniques 11, 209–212 (1997). https://doi.org/10.1023/A:1018469901326
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DOI: https://doi.org/10.1023/A:1018469901326