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

作者：Huang, Guowei; Wang, Yun; Wang, Juping; Yang, Chunzhang; Huang, Tao; Zhuang, Zhengping; Gu, Jiang^*

来源：BMC Cell Biology, 2014, 15(1): 45.

DOI：10.1186/s12860-014-0045-1

• Morphology
• Protein
• Amino acid
• Co-localization

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.

• 出版日期2014-12-5
• 单位汕头大学