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

Author:

Bugga Paramesha123,Stoner Michael W.123,Manning Janet R.123,Mushala Bellina A.S.123,Bhattarai Nisha123,Sharifi-Sanjani Maryam123,Webster Bradley R.4,Thapa Dharendra5,Scott Iain123ORCID

Affiliation:

1. 1Vascular Medicine Institute, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.

2. 2Center for Metabolism and Mitochondrial Medicine, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.

3. 3Division of Cardiology, Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.

4. 4Department of Urology, Roswell Park Cancer Center, Buffalo, NY 14263, U.S.A.

5. 5Department of Human Performance - Exercise Physiology, West Virginia University, Morgantown, WV 26506, U.S.A.

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.

Funder

HHS | NIH | National Heart, Lung, and Blood Institute

American Heart Association

HHS | NIH | National Institute of Diabetes and Digestive and Kidney Diseases

HHS | NIH | National Institute of General Medical Sciences

Publisher

Portland Press Ltd.

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