Beauregard G, Giroux S, Potier M
Anal Biochem. 1983 Jul 15;132(2):362-4. doi: 10.1016/0003-2697(83)90021-0.
Target size analysis by radiation inactivation is now a well-established method to study structure-function relationships in biologically active macromolecules without prior purification or even solubilization. Recently, it was reported that a relatively low-dose-rate but commonly available gamma source such as the Gammacell 220 (Atomic Energy of Canada, Ltd.) can be used to carry out radiation inactivation experiments providing it is appropriately calibrated with enzymes of known radiation sensitivities (G. Beauregard and M. Potier (1982) Anal. Biochem. 122, 379-384). In this report, a tube rack designed to fit into the irradiation chamber of the Gammacell 220 which allows five experiments (at 30 tubes per experiment) to be carried out simultaneously with both standard and unknown samples is described. The dose rates delivered at different positions in the rack were determined by irradiating rat liver cytosolic neuraminidase, an enzyme of known radiation sensitivity. A better than 2.7% agreement was obtained between experimental dose rate and computed values from isodose curves previously published by other authors (O. A. Curzio and H. O. Quaranta (1982) Int. J. Appl. Radiat. Isot. 33, 1-3).
通过辐射失活进行靶标大小分析,如今已成为一种成熟的方法,可用于研究生物活性大分子的结构-功能关系,而无需事先进行纯化甚至溶解。最近有报道称,一种剂量率相对较低但常见的伽马源,如伽马细胞220(加拿大原子能有限公司),如果用已知辐射敏感性的酶进行适当校准,就可用于开展辐射失活实验(G. 博勒加德和M. 波捷(1982年)《分析生物化学》122卷,第379 - 384页)。在本报告中,描述了一种设计用于适配伽马细胞220辐照室的试管架,它能同时对标准样品和未知样品进行五项实验(每项实验30个试管)。通过辐照大鼠肝脏胞质神经氨酸酶(一种已知辐射敏感性的酶),测定了试管架不同位置处的剂量率。实验剂量率与其他作者先前发表的等剂量曲线计算值之间的一致性优于2.7%(O. A. 库尔齐奥和H. O. 夸兰塔(1982年)《国际应用辐射与同位素杂志》33卷,第1 - 3页)。