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a new microsphere-based immunoassay for measuring the activity of transcription factors新microsphere-based免疫分析法测量转录因子的活性.pdf

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a new microsphere-based immunoassay for measuring the activity of transcription factors新microsphere-based免疫分析法测量转录因子的活性

Biol Proced Online (2010) 12:18–26 DOI 10.1007/s12575-010-9030-z A New Microsphere-Based Immunoassay for Measuring the Activity of Transcription Factors Yu-Ling Lin Yun-Ju Lai Nu-Man Tsai Tai-Chu Peng Yen-Ku Liu Ru-Ping Lee Chueh-Jen Tsai Kuang-Wen Liao Received: 14 September 2009 /Accepted: 16 March 2010 /Published online: 14 April 2010 # The Author(s) 2010. This article is published with open access at S Abstract There are several traditional and well-developed cultured in the wells of 96-well titer plates. Thus, a new methods for analyzing the activity of transcription factors, microsphere-based immunoassay to measure the activity of such as EMSA, enzyme-linked immunosorbent assay, and transcription factors (MIA-TF) was developed. In MIA-TF, reporter gene activity assays. All of these methods have NeutrAvidin-labeled microspheres were used as the solid their own distinct disadvantages, but none can analyze the phase to capture biotin-labeled double-strand DNA frag- changes in transcription factors in the few cells that are ments which contain certain transcription factor binding elements. The activity of transcription factors was detected by immunoassay using a transcription factor-specific Yu-Ling Lin, Yun-Ju Lai and Tai-Chu Peng contributed equally to this work. antibody to monitor the binding with the DNA probe. Next, analysis was performed by flow cytometry. The Dr. Kuang-Wen Liao and Dr. Ru-Ping Lee contributed equally to t