Development and clinical application of quantity profile analysis to inflammatory allied transcription factor activity by fluorescence correlation spectroscopy

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K08612
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory medicine
• Research Institution: University of Toyama
• Principal Investigator: Kitajima Isao 富山大学, 大学院医学薬学研究部(医学), 教授 (50214797)
• Co-Investigator(Kenkyū-buntansha): 仁井見 英樹 富山大学, 大学院医学薬学研究部(医学), 准教授 (50401865)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 転写因子 / ＮＦ－κＢ / 蛍光相関分光法 / ＥＬＩＳＡ / 活性定量

Outline of Final Research Achievements

As for the quantity transcription factor activated analysis, an ELISA method is used at a study level. We developed new experimental apparatus for the detection of inflammatory allied transcription factor activity based on fluorescence correlation spectroscopy (FCS), which is able to analyze binding between DNA and protein in the liquid phase. We made each fluorescence DNA probe which specifically bound to NF-κB, AP1, NF-IL-6, SP1, NF-AT or OCT1. To assess the applicability of the FCS method, we quantitated each transcription factor activity in the human lymphocytes stimulated with TNF-α, LPS, or PMA/Ionomycin. In results, 1.5-fold increased NF-κB and AP1 activity was measured two hours after stimulation with TNF-α or PMA/Ionomycin. The quantity transcription assay by the FCS method is quick, simple, easy and high-throughput analytical method, compared to the ELISA method.

Free Research Field

臨床検査医学