Koenig E
J Cell Biol. 1968 Sep;38(3):562-73. doi: 10.1083/jcb.38.3.562.
A method is described for the microanalysis of protein, obtained from isolated tissue elements, in the range of 500 micromicrog-500 mmicrog. The method entails solubilization of cellular protein with phosphoric acid and heat after extraction of acid-soluble compounds, lipids, and RNA. A procedure for the extraction and recovery of cellular RNA by the use of 40% trichloroacetic acid is presented. The solubilized protein, in the form of a microdroplet, is photomicrographed with monochromatic light at 230 mmicro. Total density in the microdroplet is determined from calibrated photographic plates by microdensitometry, and is converted to protein mass by using an experimentally determined average specific absorbance value. A solubilized protein labeled with tritium can be recovered after photomicrography, combusted, and reduced to generate tritiated gas for high-efficiency tritium radiometry. Total protein was analyzed in (a) nerve cells of three different sizes from Deiters' nucleus of the rabbit; and the whole rod cell and rod cell nucleus of the rabbit retina.
本文描述了一种用于对从分离的组织成分中获得的蛋白质进行微量分析的方法，分析范围为500微微克至500微克。该方法包括在提取酸溶性化合物、脂质和RNA后，用磷酸和加热使细胞蛋白溶解。本文还介绍了一种使用40%三氯乙酸提取和回收细胞RNA的方法。以微滴形式存在的溶解蛋白，在230微米处用单色光进行显微摄影。通过微量光密度测定法从校准的照相底片上确定微滴中的总密度，并使用实验确定的平均比吸光值将其转换为蛋白质量。用氚标记的溶解蛋白在显微摄影后可以回收，燃烧并还原以产生用于高效氚放射性测量的氚气。对以下样本进行了总蛋白分析：(a)来自兔Deiters核的三种不同大小的神经细胞；以及兔视网膜的整个视杆细胞和视杆细胞核。
