聚蛋白多糖酶
阿格里坎
阿达姆斯
抗体
化学
细胞生物学
免疫学
生物
医学
关节软骨
生物化学
酶
骨关节炎
金属蛋白酶
血栓反应素
病理
替代医学
作者
Salvatore Santamaria,Kazuhiro Yamamoto
出处
期刊:Methods in molecular biology
日期:2019-08-29
卷期号:: 125-136
被引量:10
标识
DOI:10.1007/978-1-4939-9698-8_11
摘要
Aggrecan is a major matrix component of articular cartilage, and its dysregulated proteolysis is a crucial event in the pathogenesis of arthritis. Aggrecanases, members of ADAMTS family, play a pivotal role in aggrecan degradation with ADAMTS-4 and ADAMTS-5 being key enzymes. Cleavage events mediated by ADAMTSs are highly specific and very well characterized; therefore, it is possible to investigate aggrecanolysis by using antibodies reacting with the new N- and C-termini of the cleavage products (neoepitope antibodies). Here, we present a method for analyzing dynamic aggrecanolysis by Western blotting using neoepitope antibodies in combination with antibodies against total aggrecan fragments. The protocol is robust and has a broad application for investigation of aggrecanase activity in vitro and ex vivo.
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