Immunofluorescent quantification of tyrosine phosphorylated proteins by flow cytometric analysis

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Title
Immunofluorescent quantification of tyrosine phosphorylated proteins by flow cytometric analysis
Author(s)
Do Young Yoon; Sun Young Yoon; Woo Suk Koh; In Seong Choe; Tai Wha Chung; Mi Young Han
Bibliographic Citation
Biotechnology Techniques, vol. 11, no. 3, pp. 209-211
Publication Year
1997
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 (30 μg/1 x 106cells). 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.
ISSN
0951-208X
Publisher
Springer
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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