Watkins R W, Robertson C R
J Biomed Mater Res. 1977 Nov;11(6):915-38. doi: 10.1002/jbm.820110611.
A total internal-reflection fluorescence (TIRF) technique for examining protein adsorption at solid-liquid interfaces is described. For the representative case of adsorption of bovine gamma-globulin onto silicone rubber, the applicability of the technique to serveral important aspects of protein adsorption phenomena is demonstrated. Specifically, in addition to the tightly adsorbed protein layer observed herein and previously by maany others, the presence of a loosely adsorbed protein layer was also noted under conditions of either static or flowing protein solutions. Taking advantage of the continuous real-time measurements possible with TIRF, a rate constant for the protein adsorption step during laminar flow was estimated at both 25 and 37 degrees C. These results serve to demonstrate that the TIRF technique represents a significant and versatile new tool for the study of protein adsorption phenomena.
描述了一种用于研究固液界面蛋白质吸附的全内反射荧光（TIRF）技术。对于牛γ-球蛋白吸附到硅橡胶上的典型情况，证明了该技术在蛋白质吸附现象的几个重要方面的适用性。具体而言，除了本文以及许多其他人之前观察到的紧密吸附的蛋白质层外，在静态或流动的蛋白质溶液条件下，还注意到存在松散吸附的蛋白质层。利用TIRF可以进行连续实时测量的优势，估算了在25℃和37℃下层流过程中蛋白质吸附步骤的速率常数。这些结果表明，TIRF技术是研究蛋白质吸附现象的一种重要且通用的新工具。
