在培养细胞和小鼠中过表达和基于 ELISA 的 asporsin 检测。
Overexpression and ELISA-based detection of asprosin in cultured cells and mice.
机构信息
Harrington Discovery Institute at University Hospitals, 2103 Cornell Road, Wolstein Research Building 4130, Cleveland, OH 44106, USA.
Harrington Discovery Institute at University Hospitals, 2103 Cornell Road, Wolstein Research Building 4130, Cleveland, OH 44106, USA; Department of Medicine, Case Western Reserve University, Cleveland, OH 44106, USA; Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, OH 44106, USA.
出版信息
STAR Protoc. 2022 Dec 16;3(4):101762. doi: 10.1016/j.xpro.2022.101762. Epub 2022 Oct 13.
The unreliability of commercial recombinant asprosin preparations and variability between asprosin detection assays have proven to be a bottleneck in experimental interpretation. This protocol describes the use of viral vectors and expression plasmid for overexpression and secretion of human asprosin to achieve sustained elevation of asprosin protein in mice and HEK293T cells without using recombinant proteins. This protocol also includes a sandwich ELISA using anti-asprosin monoclonal antibodies for detection of asprosin in media from cultured cells and in plasma of mice. For complete details on the use and execution of this protocol, please refer to Duerrschmid et al. (2017), Mishra et al. (2021), and Mishra et al. (2022).
商业重组阿普瑞素制剂的不可靠性和阿普瑞素检测分析之间的可变性已被证明是实验解释的一个瓶颈。本方案描述了使用病毒载体和表达质粒过表达和分泌人阿普瑞素，以在不使用重组蛋白的情况下实现小鼠和 HEK293T 细胞中阿普瑞素蛋白的持续升高。本方案还包括使用抗阿普瑞素单克隆抗体的夹心 ELISA 法检测培养细胞培养基和小鼠血浆中的阿普瑞素。有关本方案使用和执行的完整详细信息，请参阅 Duerrschmid 等人（2017 年）、Mishra 等人（2021 年）和 Mishra 等人（2022 年）。
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