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Title 

Immunofluorescent quantification of tyrosine phosphorylated proteins by flow cytometric analysis

Authors 

Do Young YoonSun Young YoonWoo Suk KohIn Seong ChoeTai Wha ChungMi Young Han

Publisher 

Springer

Issue Date 

1997

Citation 

Biotechnology Techniques, vol. 11, no. 3, pp. 209-211

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

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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