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

BMC Cell Biology

Methodology Article

Juping Wang¹  Tao Huang¹  Jiang Gu¹  Yun Wang¹  Guowei Huang¹  Chunzhang Yang²  Zhengping Zhuang² 

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

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

DOI  :  10.1186/s12860-014-0045-1

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

来源: Springer

【 摘 要 】

BackgroundWith 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.ResultsIn 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.ConclusionsThis 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】

Unknown   
© Huang et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.