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目的 :探讨有效提取膜迷路蛋白时十二烷基硫酸钠 (SDS)的用量。方法 :用二喹啉甲酸(BCA)法测定含不同浓度SDS的提取液所提取的豚鼠内耳膜迷路蛋白含量 ,并用SDS 聚丙烯酰胺凝胶电泳 (PAGE)分析其蛋白组分。结果 :0 .2 3%SDS、1%SDS、2 %SDS组所提取的蛋白量大于 0 .1%SDS组 ,同时电泳显示的条带更多 ,染色更深。 1%SDS组与 2 %SDS组效果又好于0 .2 3%SDS组 ,其中 2 %SDS组提取的量与 1%SDS组差别不大 ,但去垢剂 蛋白质之比明显升高。结论 :蛋白提取液中含 1%SDS ,去垢剂 蛋白质比为 5～ 6左右时对充分提取膜迷路蛋白较为合适。
Objective: To study the dosage of sodium dodecyl sulfate (SDS) for the effective extraction of membrane laccase. Methods: The bicuculline carboxylic acid (BCA) method was used to determine the content of labyrinthine protein in the eardrum of guinea pigs extracted with extracts containing different concentrations of SDS, and its protein components were analyzed by SDS polyacrylamide gel electrophoresis (PAGE). Results: 0.2% SDS, 1% SDS, 2% SDS extracted more protein than 0.1% SDS group, meanwhile electrophoresis showed more bands and deeper staining. The effect of 1% SDS group and 2% SDS group was better than that of 0.2% SDS group, but the difference between 2% SDS group and 1% SDS group was not significant, but the ratio of detergent protein was significantly increased. CONCLUSION: It is more appropriate to fully extract the laccase when the protein extract contains 1% SDS and the detergent protein ratio is about 5 ~ 6.