FILM DETECTION METHOD FOR TRITIUM-LABELED PROTEINS AND NUCLEIC-ACIDS IN POLYACRYLAMIDE GELS
FILM DETECTION METHOD FOR TRITIUM-LABELED PROTEINS AND NUCLEIC-ACIDS IN POLYACRYLAMIDE GELS
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DOI：
10.1111/j.1432-1033.1974.tb03599.x
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发表时间：
1974-01-01
期刊：
影响因子：
--
通讯作者：
LASKEY, RA
中科院分区：
文献类型：
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作者：
BONNER, WM;LASKEY, RA
本文介绍了一种用X射线胶片闪烁自显影（荧光照相）检测聚丙烯酰胺凝胶中3 H的简单方法。将凝胶在二甲亚砜中脱水，浸泡在2，5-二苯基恶唑（PPO）的二甲亚砜溶液中，干燥并在-70 °C下暴露于RP皇家“X‐Omat”膜。描述了每一步的最佳条件。来自3 H的β粒子与2，5-二苯基恶唑相互作用，发光，导致X射线胶片局部变黑。产生的图像类似于由具有较高发射能量的同位素（如14 C）的常规放射自显影获得的图像。在24 h暴露中，可检测到凝胶条带中的3000 dis.3H/min。同样，500分/用于聚丙烯酰胺凝胶中~（35）S和~（14）C的测定，比常规放射自显影法灵敏10倍以上。在24小时内可检测到凝胶中一条带中的130 dis.35S或14 C/min。
A simple method is described for detecting3H in polyacrylamide gels by scintillation autography (fluorography) using X‐ray film. The gel is dehydrated in dimethyl sulphoxide, soaked in a solution of 2,5‐diphenyloxazole (PPO) in dimethylsulphoxide, dried and exposed to RP Royal “X‐Omat” film at ‐70 °C. Optimal conditions for each step are described. β‐particles from3H interact with the 2,5‐diphenyloxazole emitting light which causes local blackening of an X‐ray film. The image produced resembles that obtained by conventional autoradiography of isotopes with higher emission energies such as14C. 3000 dis.3H/min in a band in a gel can be detected in a 24‐h exposure. Similarly 500 dis./min can be detected in one week.When applied to the detection of35S and14C in polyacrylamide gels, this method is ten times more sensitive than conventional autoradiography. 130 dis.35S or14C/min in a band in a gel can be detected in 24 h.