Separation of Histone by Capillary Electrophoresis with Ultraviolet-Visible Detection

• Main Authors: LAI, YU-JHEN, 賴芋蓁
• Other Authors: CHANG,PO-LING
• Format: Others
• Language: zh-TW
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/t8u3bh

碩士 === 東海大學 === 化學系 === 105 === Histones are the basic structural units protein found in eukaryotic cell.They are highly basic amino acids which can interact with negatively charged phosphate groups in DNA. They are the chief protein components of chromatin, acting as spools around which DNA winds, and playing a role in gene regulation. Heterogeneity among cancer cells within individual tumors has emerged as a general feature of cancer,with critical implications for cancer diagnosis and treatment. That demonstrate a critical role for Histone H1.0 in inhibiting tumor maintenance. Here, we present standard protocols for acid extraction of histones from human liver cancer cells. Separation of five major families of histones include H1/H5, H2A, H2B, H3, and H4 with ultraviolet visible detection,and the detection wavelength is 280 nm. The separation condition of histone is Tris-Borate buffer add SDS (Sodium dodecyl sulfate),and the separation voltage is +15 kV. By changing the buffer solution concentration, pH value, and SDS concentration can improve the protein will be adsorbed on the surface of the capillary. By adding different concentrations of 250K Da hydroxyethyl cellulose, the best separation resolution can be obtained. Under through optimized experimental conditions and apparatus , it can separate the different kind of cancer cell include Nasopharyngeal carcinoma cells (TW02), cervical cancer cells (Hela), lung cancer cells (A549). We can explore the differences in histones between different cancer cells.