①测定过程中试剂二、试剂三和粗酶液均需置于冰上放置,以免变性或失活;
②若A1测定大于1.5或∆A大于0.3,建议将粗酶液使用蒸馏水适当稀释后再进行测定;若∆A小于0.02,建议适当延长酶促反应时间或增加上样量后再进行测定,计算时相应修改;
③准确在20 s和320 s处完成吸光值测定,以保证实验结果的准确性和重复性;若同时测定多个样本时,应使用多道移液器且分批进行测定,以确保组间反应时间一致;
④提取液A中含有约1 mg/mL的蛋白,测定粗酶液蛋白浓度时需减去提取液A自身的蛋白含量;
⑤沸水浴处理过程推荐使用螺纹盖离心管或冻存管,以确保密封效果防止水分挥发;
[1] Zhang Y, Gao Y, Wang J, et al. Bioenergetic Metabolism Modulatory Peptide Hydrogel for Cardiac Protection and Repair After Myocardial Infarction[J]. Advanced Functional Materials, 2024: 2312772.(IF 19)
[2] Shi L, Zhao X, Zhong K, et al. Physiological mechanism of the response to Cr (VI) in the aerobic denitrifying ectomycorrhizal fungus Pisolithus sp. 1[J]. Journal of Hazardous Materials, 2022, 429: 128318.(IF 14.224)
[3] Deng L, Liu M, Yu Y, et al. Amino acid mutation of succinate dehydrogenase complex induced resistance to benzovindiflupyr in Magnaporthe oryzae[J]. Pesticide Biochemistry and Physiology, 2024, 203: 106027. (IF 4.2)
[4] Zhu P, Wang P, Teng Q, et al. Postharvest Preservation of Flammulina velutipes with Isoamyl Isothiocyanate[J]. Agronomy, 2023, 13(7): 1771.(IF 3.7)