Department of Biological and Biomedical Sciences, Rowan University, 201 Mullica Hill Rd, Glassboro, NJ, 08028, USA.
Department of Chemistry and Biochemistry, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA, 92093-0358, USA.
Sci Rep. 2023 Nov 22;13(1):20529. doi: 10.1038/s41598-023-46571-5.
Accurate protein quantitation is essential for many cellular mechanistic studies. Existing technology relies on extrinsic sample evaluation that requires significant volumes of sample as well as addition of assay-specific reagents and importantly, is a terminal analysis. This study exploits the unique chemical features of a fluorescent molecular rotor that fluctuates between twisted-to-untwisted states, with a subsequent intensity increase in fluorescence depending on environmental conditions (e.g., viscosity). Here we report the development of a rapid, sensitive in situ protein quantitation method using ARCAM-1, a representative fluorescent molecular rotor that can be employed in both non-terminal and terminal assays.
准确的蛋白质定量对于许多细胞机制研究至关重要。现有的技术依赖于外在的样本评估,这需要大量的样本以及添加特定于检测的试剂,而且重要的是,这是一种末端分析。本研究利用荧光分子转子的独特化学特性,该分子转子在扭曲到未扭曲状态之间波动,随后荧光强度根据环境条件(例如,粘度)增加。在这里,我们报告了一种使用 ARCAM-1 的快速、灵敏的原位蛋白质定量方法的开发,该分子转子是一种代表性的荧光分子转子,可用于非末端和末端检测。