Falk B W, Elliott C
Anal Biochem. 1985 Feb 1;144(2):537-41. doi: 10.1016/0003-2697(85)90151-4.
Fluorescent labeling of proteins was found to be a very sensitive and reliable alternative to conventional methods for monitoring proteins on Western blots. Proteins were labeled with 2-methoxy-2,4-diphenyl-3(2H)-furanone (MDPF) before SDS-PAGE. After electrophoresis and subsequent electro-blotting the fluorescent-labeled proteins were visible upon ultraviolet illumination of the nitrocellulose membranes, and could be photographed to yield an accurate record of the blots before subsequent serological analysis. The sensitivity for detecting MDPF-labeled proteins on nitrocellulose was 100-200 ng, 50 to 100 fold less sensitive than on gels. Fluorescent-labeled TMV and MStpV capsid proteins that were blotted onto nitrocellulose still reacted in serological tests and were detected when present in quantities as low as 100 pg. Fluorescent labeling allows accurate photographic records of the SDS-gel, blot and probed blot using only one sample, and no subsequent staining steps are required.
蛋白质的荧光标记被发现是一种非常灵敏且可靠的替代传统方法，用于在蛋白质印迹法中监测蛋白质。在进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）之前，用2-甲氧基-2,4-二苯基-3(2H)-呋喃酮（MDPF）标记蛋白质。电泳及随后的电印迹之后，在硝酸纤维素膜经紫外线照射时，荧光标记的蛋白质可见，并且在后续血清学分析之前可拍照以获得印迹的准确记录。在硝酸纤维素膜上检测MDPF标记蛋白质的灵敏度为100 - 200纳克，比在凝胶上的灵敏度低50至100倍。印迹到硝酸纤维素膜上的荧光标记烟草花叶病毒（TMV）和马铃薯X病毒（MStpV）衣壳蛋白在血清学试验中仍有反应，当含量低至100皮克时也能被检测到。荧光标记仅使用一个样品就能对SDS凝胶、印迹和经探针检测的印迹进行准确的拍照记录，并且无需后续染色步骤。
