1. A morphologic and semi-quantitative technique to analyze synthesis and release of specific proteins in cells.
- Author
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Guowei Huang, Yun Wang, Juping Wang, Chunzhang Yang, Tao Huang, Zhengping Zhuang, and Jiang Gu
- Subjects
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PROTEIN synthesis , *CELL morphology , *IMMUNOGLOBULIN G , *CELL culture , *ENZYME-linked immunosorbent assay - Abstract
Background: With the rapid advancement of cell biology, the evaluation of a given protein's synthesis and release in cells becomes critical. However, up to now there has been no technique available to morphologically visualize and measure a newly synthesized protein in cells, nor can we measure the protein's release from the cells. Results: In this study, we developed a set of assays combining pulse chase amino acid substitution, non-radioactive labeling, and immunofluorescence co-localization to visualize newly synthesized proteins in individual cells and then to detect their release using modified ELISA. We demonstrated the synthesis and release of Bcl-2, MMP-9, and immunoglobulin G (IgG) in a human trophoblast cell line, of which the last finding has not been reported previously. Conclusions: This new technique offers a powerful tool to evaluate the dynamics of the synthesis and release of target proteins in individual cultured cells with wide applications in genetic and protein analysis. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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