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市售的PGC-1α抗体在特异性和灵敏度方面差异很大。

Commercially available PGC-1α antibodies vary greatly in specificity and sensitivity.

作者信息

Galipeau Maria, Schmitt Clémence, Baldwin Cindy, Gunes Aysim, Estall Jennifer L

机构信息

Faculty of Medicine, University of Montreal, Montreal, Quebec, Canada.

Institut de Recherches Cliniques de Montréal, Montreal, Quebec, Canada.

出版信息

MicroPubl Biol. 2024 Aug 23;2024. doi: 10.17912/micropub.biology.001268. eCollection 2024.

Abstract

Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) is an inducible transcriptional coactivator protein involved in mitochondrial biogenesis and metabolism. PGC-1α exhibits a short half-life and low abundance, rendering its detection challenging by immunoblotting. This study compared the specificity and sensitivity of seven commercially available PGC-1α antibodies towards the full-length mouse PGC-1α1 isoform, using overexpression and knockdown in primary mouse hepatocytes. While all antibodies identified overexpressed PGC-1α1, only one detected endogenous PGC-1α1. This study demonstrates wide variability in sensitivity and specificity of PGC-1α antibodies and suggests that controls should be used to differentiate PGC-1α protein from non-specific bands.

摘要

过氧化物酶体增殖物激活受体γ共激活因子1α(PGC-1α)是一种可诱导的转录共激活蛋白,参与线粒体生物合成和代谢。PGC-1α半衰期短且丰度低,通过免疫印迹法检测具有挑战性。本研究使用原代小鼠肝细胞中的过表达和敲低方法,比较了七种市售PGC-1α抗体对全长小鼠PGC-1α1亚型的特异性和敏感性。虽然所有抗体都能识别过表达的PGC-1α1,但只有一种能检测到内源性PGC-1α1。本研究表明PGC-1α抗体的敏感性和特异性存在很大差异,并建议应使用对照来区分PGC-1α蛋白与非特异性条带。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f3b9/11380096/9ecb5edeab8f/25789430-2024-micropub.biology.001268.jpg

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