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目的建立一种冬虫夏草蛋白质含量快速测定方法,弥补Bradford法不足,为研究过程中快速准确定量蛋白质奠定基础。方法本研究用牛血清蛋白(bovine serum albumin,BSA)作为已知浓度的定量标准,用它来计算其它带的量。根据Quantity One来降低图像噪音和背景强度,并确定泳道,定义带并建立光密度与已知浓度的定量标准蛋白线性关系,通过每个带的光密度定量计算凝胶上每个带的质量。结果本实验所建立的凝胶染色扫描法能够较为快速、简便、准确地测定冬虫夏草蛋白含量。结论凝胶染色扫描法用于蛋白质定量,准确度、精密度、重复性均符合要求,为测定方法的进一步完善奠定了基础。
OBJECTIVE To establish a rapid method for the determination of protein content in Cordyceps sinensis, to make up for the deficiencies of Bradford method and to lay a foundation for the rapid and accurate determination of protein during the research. Methods In this study, bovine serum albumin (BSA) was used as a quantitative standard of known concentration and used to calculate the amount of other bands. Decrease image noise and background intensity based on Quantity One, and determine lanes, define bands, and establish a linear relationship between optical density and a quantified standard protein of known concentration. The mass of each band on the gel is quantified by the optical density of each band. Results The gel staining scanning method established in this experiment can determine the protein content of Cordyceps more quickly, easily and accurately. Conclusion The method of gel staining scanning for protein quantification, accuracy, precision and reproducibility all meet the requirements, which lays the foundation for the further improvement of the determination method.