双线传奇私服-新抗体可用于自噬诱导的分析

时间:2019-11-27 15:37 来源: www.seosem.ws

本期文章:《自然—方法学》:Online/在线发表

加拿大渥太华大学Ryan C. Russell研究组报道了一种能够用于分析自噬产生的抗体。2019年11月25日,国际知名学术期刊《自然—方法学》在线发表了这一成果。

研究人员报道了一种通过测量蛋白ATG16L1的磷酸化来监测自噬的方法。研究人员开发并鉴定了可以在哺乳动物细胞中内源性检测磷酸化ATG16L1的单克隆抗体。

重要的是,磷酸化ATG16L1仅存在于新形成的自噬体上。因此,它的水平不受长时间的压力或后期自噬阻滞的影响,这两个因素会混淆自噬分析。

此外,研究人员表明ATG16L1磷酸化是由许多自噬诱导应激源激活的保守的信号通路。这一抗体适用于蛋白质印迹、免疫荧光和免疫组化,并且测得的磷酸化ATG16L1水平直接对应于自噬速率。

总之,这个磷酸抗体是一种研究自噬诱导的新工具。

据介绍,自噬是一种可维持细胞稳态的降解程序。自噬缺陷已在多种疾病中有报道。然而,由于目前可用的自噬诱导测量工具的局限性,自噬速率的分析可能具有挑战性,特别是在稀有细胞群体或体内。

附:英文原文

Title: An antibody for analysis of autophagy induction

Author: Wensheng Tian, Reham Alsaadi, Zhihao Guo, Alena Kalinina, Mical Carrier, Marie-Eve Tremblay, Baptiste Lacoste, Diane Lagace, Ryan C. Russell

Issue&Volume: 2019-11-25

Abstract: Autophagy is a degradative program that maintains cellular homeostasis. Autophagy defects have been described in numerous diseases. However, analysis of autophagy rates can be challenging, particularly in rare cell populations or in vivo, due to limitations in currently available tools for measuring autophagy induction. Here, we describe a method to monitor autophagy by measuring phosphorylation of the protein ATG16L1. We developed and characterized a monoclonal antibody that can detect phospho-ATG16L1 endogenously in mammalian cells. Importantly, phospho-ATG16L1 is only present on newly forming autophagosomes. Therefore, its levels are not affected by prolonged stress or late-stage autophagy blocks, which can confound autophagy analysis. Moreover, we show that ATG16L1 phosphorylation is a conserved signaling pathway activated by numerous autophagy-inducing stressors. The described antibody is suitable for western blot, immunofluorescence and immunohistochemistry, and measured phospho-ATG16L1 levels directly correspond to autophagy rates. Taken together, this phospho-antibody represents an exciting tool to study autophagy induction.

DOI: 10.1038/s41592-019-0661-y

Source: https://www.nature.com/articles/s41592-019-0661-y

期刊信息

Nature Methods:《自然—方法学》,创刊于2004年。隶属于施普林格·自然出版集团,最新IF:28.467
官方网址:https://www.nature.com/nmeth/
投稿链接:https://mts-nmeth.nature.com/cgi-bin/main.plex