Cawood A H, Potter U, Dickinson H G
J Histochem Cytochem. 1978 Aug;26(8):645-50. doi: 10.1177/26.8.80420.
The specificity and stoichiometry of the binding of Coomassie Brilliant Blue (CBB) to protein in section has been examined using both frozen protein matrices and plant material. The maximum adsorbance of the stain, bound and in solution, was found to be 620 nm although variation in the results at this wavelength necessitated measurements to be made at 600 nm. After enzyme treatments of sectioned plant material embedded in resin, all CBB-binding biological material was shown to be sensitive to non-specific protease. The relationship between optical density at 600 nm and section thickness was tested statistically against the Lambert-Beer law, using microdensitometry of cryostat-sectioned, frozen genatine solution. The analyses showed conclusively that, under these conditions, CBB adheres strongly to the Lambert-Beer relationship. CBB may thus be considered as a very specific protein stain, eminently suited both to cytological observation and quantitative microdensitometry.
使用冷冻蛋白质基质和植物材料，研究了考马斯亮蓝（CBB）与切片中蛋白质结合的特异性和化学计量关系。尽管在此波长下结果存在差异，需要在600nm处进行测量，但发现结合态和溶液态染色剂的最大吸光度为620nm。对包埋在树脂中的植物材料切片进行酶处理后，所有与CBB结合的生物材料均显示对非特异性蛋白酶敏感。使用低温恒温器切片的冷冻明胶溶液的显微密度测定法，根据朗伯-比尔定律对600nm处的光密度与切片厚度之间的关系进行了统计学测试。分析结果明确表明，在这些条件下，CBB与朗伯-比尔关系紧密相关。因此，CBB可被视为一种非常特异的蛋白质染色剂，非常适合细胞学观察和定量显微密度测定。
