Gubler M L, Abarzúa P
Department of Oncology, Roche Research Center, Hoffmann-LaRoche Inc., Nutley, NJ 07110, USA.
Biotechniques. 1995 Jun;18(6):1008, 1011-4.
A nonradioactive functional assay was developed to quantitate DNA binding proteins. The assay was designed to allow the use of 96-well microplates for high sample throughput. We show that the assay can measure sequence-specific DNA binding of purified proteins as well as DNA binding activity present in whole cell extracts. By using a temperature-sensitive DNA binding protein, we demonstrate that DNA binding as measured by this novel assay correlates with the biological function of the protein.
已开发出一种非放射性功能测定法来定量DNA结合蛋白。该测定法旨在允许使用96孔微孔板以实现高样本通量。我们表明,该测定法可以测量纯化蛋白的序列特异性DNA结合以及全细胞提取物中存在的DNA结合活性。通过使用一种温度敏感的DNA结合蛋白,我们证明通过这种新测定法测量的DNA结合与该蛋白的生物学功能相关。