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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。
In this paper, the toxin protein in Bacillus thuringiensis preparation by SDS-PAGE-elution colorimetric analysis method. B.t. samples were treated with NaOH solution, followed by L aemmli buffer treatment, centrifugation, the supernatant was electrophoresis. After staining with Coomassie brilliant blue (CBB) R-2 50, the 130 KD-toxin protein band was scraped off, the protein-adsorbed CBB R-2 50 was eluted with pyridine, the absorbance of the solution was measured at 650 nm, Quantitative. The method of high accuracy, good precision. RSD at about 5%. The recoveries ranged from 94.6% to 99.2%. This method is not significantly different from the result of SDS-PAGE-TL C-scan. There is also a good correlation between the amount of 130 KD-toxin protein and the bioassay titer.