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Validation of GCN5L1/BLOC1S1/BLOS1 antibodies using knockout cells and tissue.
Bugga, Paramesha; Stoner, Michael W; Manning, Janet R; Mushala, Bellina A S; Bhattarai, Nisha; Sharifi-Sanjani, Maryam; Webster, Bradley R; Thapa, Dharendra; Scott, Iain.
Affiliation
  • Bugga P; Vascular Medicine Institute, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.
  • Stoner MW; Center for Metabolism and Mitochondrial Medicine, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.
  • Manning JR; Division of Cardiology, Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.
  • Mushala BAS; Vascular Medicine Institute, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.
  • Bhattarai N; Center for Metabolism and Mitochondrial Medicine, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.
  • Sharifi-Sanjani M; Division of Cardiology, Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.
  • Webster BR; Vascular Medicine Institute, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.
  • Thapa D; Center for Metabolism and Mitochondrial Medicine, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.
  • Scott I; Division of Cardiology, Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, U.S.A.
Biochem J ; 481(10): 643-651, 2024 May 22.
Article de En | MEDLINE | ID: mdl-38683688
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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Texte intégral: 1 Collection: 01-internacional Base de données: MEDLINE Sujet principal: Anticorps Limites: Animals Langue: En Journal: Biochem J Année: 2024 Type de document: Article Pays d'affiliation: États-Unis d'Amérique Pays de publication: Royaume-Uni

Texte intégral: 1 Collection: 01-internacional Base de données: MEDLINE Sujet principal: Anticorps Limites: Animals Langue: En Journal: Biochem J Année: 2024 Type de document: Article Pays d'affiliation: États-Unis d'Amérique Pays de publication: Royaume-Uni