Leong L M, Tan B H, Ho K K
Department of Botany, Faculty of Science, National University of Singapore.
Anal Biochem. 1992 Dec;207(2):317-20. doi: 10.1016/0003-2697(92)90018-3.
Nonheme iron proteins can be visualized as blue bands in native polyacrylamide gels using a staining method that is both simple and rapid. The reaction of potassium ferricyanide with protein-bound iron atoms to form royal blue complexes occurs almost instantaneously and is sensitive enough to detect 1 microgram of analytical-grade ferritin and 2 micrograms of purified ferredoxin from cyanobacteria. No special treatment of reagents or apparatus was necessary. On comparison, this stain was found to be more specific than the Ferene S stain, not detecting bovine serum albumin even when present as a hundredfold excess over ferritin. The method was found to be effective for isoelectric focusing gels as well.
使用一种简单快速的染色方法,非血红素铁蛋白在天然聚丙烯酰胺凝胶中可呈现为蓝色条带。铁氰化钾与蛋白质结合的铁原子反应形成品蓝色络合物,该反应几乎瞬间发生,灵敏度足以检测到1微克分析级铁蛋白和2微克来自蓝细菌的纯化铁氧化还原蛋白。无需对试剂或仪器进行特殊处理。相比之下,发现这种染色比Ferene S染色更具特异性,即使牛血清白蛋白的含量比铁蛋白高出百倍也检测不到。该方法对等电聚焦凝胶也有效。