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

BMC Cell Biology

Jiang Gu²  Zhengping Zhuang¹  Tao Huang²  Chunzhang Yang¹  Juping Wang²  Yun Wang²  Guowei Huang² 

[1] Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of health, Bethesda 20892, Maryland, USA;Department of Pathology and Pathophysiology, Guangdong Provincial Key Laboratory of Infectious Disease and Immunopathology, Shantou University Medical College, Shantou 515041, Guangdong, China

关键词: Co-localization;    Amino acid;    Protein;    Morphology;   

Others  :  1088699
DOI  :  10.1186/s12860-014-0045-1

 received in 2014-07-15, accepted in 2014-11-25,  发布年份 2014

【 摘 要 】

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.

【 授权许可】

   
2014 Huang et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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