不同浓度十二烷基磺酸钠在提取甲醛固定石蜡包埋组织β-淀粉样前体蛋白剪切酶-1中的作用

β-myloid precursor protein cleavage enzyme-1 extracted from formalin-fixed,paraffin-embedded tissue following different sodium dodecyl sulfate concentration treatment

目的:探索从甲醛固定、石蜡包埋(formaldehyde-fixed,paraffin-embedded,FFPE)的脑组织中提取β-淀粉样前体蛋白剪切酶-1(BACE-1)的有效方法。方法:将大鼠FFPE脑组织分别加入不同浓度十二烷基磺酸钠(SDS,0%,0.5%,1%,2%和4%)提取BACE-1,新鲜大鼠脑组织作为对照组。提取的BACE-1蛋白量采用Western blot进行检测。结果:未用SDS处理的固定组织提取不到BACE-1,随着SDS浓度的逐渐升高,固定组织中提取的BACE-1蛋白也逐渐增加,4%SDS处理的FFPE组织与新鲜组织提取的BACE-1蛋白量没有明显差别。结论:SDS处理是FFPE组织提取BACE-1的必需条件,高浓度(4%)SDS处理是FFPE组织提取BACE-1的一种有效方法。

Objective： To investigate the effective melhod of β-myloid precursor protein cleavage enzyme-1 （BACE-1） extracted from formaldehyde-fixed, paraffin-embedded （FFPE） rat brain tissue. Methods： FFPE tissue was treated with sodium dodecyl sulfate （SDS） at different concentrations （0%, 0. 5 %, 1%, 2% and 4%） to extract BACE-1 protein. Fresh tissue was served as control. The BACE-1 extraction was detected by Western blot. Results： No BACE1 was detected in no SDS treated group, and the quantity of BACE-1 was increased as the concentration of SDS rose gradually. There was no significant difference between the fresh tissue and FFPE tissue which treated by 4% SDS. Conclusion： SDS is a critical chemical component in the BACE-1 extracted from FFPE tissue. The treatment of 4% SDS is an effective method to extract BACE-1 protein from FFPE tissue.