A morphologic and semi-quantitative technique to analyze synthesis and release of specific proteins in cells

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• 作者：Guowei Huang ; Yun Wang ; Juping Wang ; Chunzhang Yang ; Tao Huang…
• 关键词：Morphology ; Protein ; Amino acid ; Co ; localization
• 刊名：BMC Cell Biology
• 出版年：2014
• 出版时间：December 2014
• 年：2014
• 卷：15
• 期：1
• 全文大小：2,952 KB
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• 刊物主题：Cell Biology; Biological Microscopy; Life Sciences, general;
• 出版者：BioMed Central
• ISSN：1471-2121

文摘

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.