Stephano J L, Gould M, Rojas-Galicia L
Anal Biochem. 1986 Feb 1;152(2):308-13. doi: 10.1016/0003-2697(86)90414-8.
When acetic acid-urea polyacrylamide gels with or without Triton X-100 were immersed in 0.1 M Na picrate, pH 7, to which 1/4 vol Coomassie blue staining solution (0.2% in 45% methanol, 10% acetic acid, 45% water) was added, proteins stained rapidly (within a few minutes in gels without Triton and within an hour in gels with Triton) with little or no background staining. Thus protein bands could be observed in a single step with no destaining. The picrate-Coomassie blue method fixed and stained a small peptide (bradykinin, nine amino acids) that was not observed in gels stained with fast green, silver, or Coomassie blue following fixation in 50% trichloroacetic acid. The picrate-Coomassie blue method gave high-contrast bands suitable for densitometry. Gels containing sodium dodecyl sulfate were also stained by the picrate-Coomassie blue method if they were first washed briefly (1 h) in 45% methanol, 10% acetic acid, 45% water, presumably to remove the detergent. These gels also stained rapidly with almost no background.
将含有或不含 Triton X - 100 的醋酸 - 尿素聚丙烯酰胺凝胶浸入 pH 7 的 0.1 M 苦味酸钠溶液中，向该溶液中加入 1/4 体积的考马斯亮蓝染色液（45%甲醇、10%乙酸、45%水的混合液中含 0.2%考马斯亮蓝），蛋白质能快速染色（不含 Triton 的凝胶在几分钟内，含 Triton 的凝胶在一小时内），背景染色很少或几乎没有。因此，无需脱色就能一步观察到蛋白条带。苦味酸盐 - 考马斯亮蓝法能固定并染色一种小肽（缓激肽，九个氨基酸），而在 50%三氯乙酸固定后用固绿、银染或考马斯亮蓝染色的凝胶中未观察到该小肽。苦味酸盐 - 考马斯亮蓝法能产生适合光密度测定的高对比度条带。如果首先在 45%甲醇、10%乙酸、45%水的混合液中短暂洗涤（1 小时），含有十二烷基硫酸钠的凝胶也能用苦味酸盐 - 考马斯亮蓝法染色，推测这样做是为了去除去污剂。这些凝胶也能快速染色，几乎没有背景。
