Comprehensive Pneumology Center (CPC), University Hospital of the Ludwig-Maximilians-University (LMU) and Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), Max-Lebsche Platz 31, 81377 Munich, Germany.
STAR Protoc. 2021 May 4;2(2):100526. doi: 10.1016/j.xpro.2021.100526. eCollection 2021 Jun 18.
This protocol describes an easy and reliable in-gel proteasome assay to quantify the activity and composition of different proteasome complexes in cells and tissues. The assay works well with limited amounts of total cell protein lysates. Although this assay is optimized specifically for the proteasome chymotrypsin-like activity, it can be expanded to other proteasome activities as well. Using antibodies that detect distinct proteasome subunits or regulators, we can determine the composition and relative quantity of active proteasome complexes. For complete details on the use and execution of this protocol, please refer to Meul et al. (2020).
本方案描述了一种简便可靠的凝胶内蛋白酶体分析方法，可用于定量检测细胞和组织中不同蛋白酶体复合物的活性和组成。该方法适用于少量总细胞蛋白裂解物。尽管该测定方法是专门针对蛋白酶体糜蛋白酶样活性进行优化的，但也可以扩展到其他蛋白酶体活性。使用检测不同蛋白酶体亚基或调节剂的抗体，我们可以确定活性蛋白酶体复合物的组成和相对数量。有关该方案使用和执行的完整详细信息，请参见 Meul 等人。（2020 年）。
