Validation of GCN5L1/BLOC1S1/BLOS1 antibodies using knockout cells and tissue.

IF 4.4 3区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Biochemical Journal Pub Date : 2024-05-22 DOI:10.1042/BCJ20230302
Paramesha Bugga, Michael W Stoner, Janet R Manning, Bellina A S Mushala, Nisha Bhattarai, Maryam Sharifi-Sanjani, Bradley R Webster, Dharendra Thapa, Iain Scott
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Abstract

GCN5L1, also known as BLOC1S1 and BLOS1, is a small intracellular protein involved in many key biological processes. Over the last decade, GCN5L1 has been implicated in the regulation of protein lysine acetylation, energy metabolism, endo-lysosomal function, and cellular immune pathways. An increasing number of published papers have used commercially-available reagents to interrogate GCN5L1 function. However, in many cases these reagents have not been rigorously validated, leading to potentially misleading results. In this report we tested several commercially-available antibodies for GCN5L1, and found that two-thirds of those available did not unambiguously detect the protein by western blot in cultured mouse cells or ex vivo liver tissue. These data suggest that previously published studies which used these unverified antibodies to measure GCN5L1 protein abundance, in the absence of other independent methods of corroboration, should be interpreted with appropriate caution.

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利用基因敲除细胞和组织验证 GCN5L1/BLOC1S1/BLOS1 抗体。
GCN5L1又称BLOC1S1和BLOS1,是一种细胞内小蛋白,参与了许多关键的生物过程。在过去十年中,GCN5L1 与蛋白质赖氨酸乙酰化、能量代谢、内溶酶体功能和细胞免疫途径的调控有关。越来越多已发表的论文使用市售试剂来检测 GCN5L1 的功能。然而,在许多情况下,这些试剂并没有经过严格的验证,可能会导致误导性的结果。在本报告中,我们测试了几种市售的 GCN5L1 抗体,发现三分之二的抗体在培养的小鼠细胞或体内外肝脏组织中不能通过 Western 印迹明确检测到该蛋白。这些数据表明,在没有其他独立方法证实的情况下,以前发表的使用这些未经验证的抗体来测量 GCN5L1 蛋白丰度的研究应该谨慎解读。
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来源期刊
Biochemical Journal
Biochemical Journal 生物-生化与分子生物学
CiteScore
8.00
自引率
0.00%
发文量
255
审稿时长
1 months
期刊介绍: Exploring the molecular mechanisms that underpin key biological processes, the Biochemical Journal is a leading bioscience journal publishing high-impact scientific research papers and reviews on the latest advances and new mechanistic concepts in the fields of biochemistry, cellular biosciences and molecular biology. The Journal and its Editorial Board are committed to publishing work that provides a significant advance to current understanding or mechanistic insights; studies that go beyond observational work using in vitro and/or in vivo approaches are welcomed. Painless publishing: All papers undergo a rigorous peer review process; however, the Editorial Board is committed to ensuring that, if revisions are recommended, extra experiments not necessary to the paper will not be asked for. Areas covered in the journal include: Cell biology Chemical biology Energy processes Gene expression and regulation Mechanisms of disease Metabolism Molecular structure and function Plant biology Signalling
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