摘要
本文研究了苏云金杆菌制剂中毒素蛋白的 SDS- PAGE-洗脱比色定量分析方法。B.t.样本用 Na OH溶液处理 ,然后加 L aemmli缓冲液处理 ,离心 ,取上清液进行电泳。用考马斯亮兰 (CBB) R- 2 50染色后 ,刮下130 KD-毒素蛋白区带 ,用吡啶洗脱蛋白吸附的 CBB R- 2 50 ,于 6 0 5nm测定溶液的吸光度 ,间接对毒素蛋白进行定量。本方法准确度高、精密度好。 RSD在 5%左右。添加回收率介于 94 .6 %～ 99.2 %。本方法与 SDS-PAGE- TL C-扫描法结果相比没有显著差异。 130 KD-毒素蛋白的量与生物测定得到的效价之间也具有较好的相关性。
The quantitative method, Sodium Dodecyl Sulphate Polyacryamide Gel Electrophoresis (SDS-PAGE)-Colorimetry, of toxin proteins in various -B.t.- commercial products was presented. -B.t.- samples were pretreated with aqueous NaOH at high pH, and treated with Laemmli sample buffer, then electrophoresis was carried out. After stained with Coomassie Brilliant Blue (CBB) R-250, the band of 130KD-toxin proteins was cut out, and the absorbed CBB R-250 was eluted with pyridine. The absorbance of the eluted dye was measured at 605nm. This method characterized by relatively high accuracy and precision. Relative standard deviation (RSD) was nearly 5%. The fortified recoveries varied from 94.6%～99.2%. There were no significant differences between the data from SDS-PAGE-Colorimetry and SDS-PAGE-TLC-Scanner. The relationship between the percentage of 130KD-toxin protein and the potency of insect bioassay was studied.
出处
《农药学学报》
CAS
CSCD
1999年第2期73-81,共9页
Chinese Journal of Pesticide Science
