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LSA-50 paper: An alternative to P81 phosphocellulose paper for radiometric protein kinase assays.
Appelmans, Olivia; Kashyap, Rudra Shankar; Gilles, Philippe; De Borggraeve, Wim M; Voet, Arnout; Van Lint, Johan.
Afiliação
  • Appelmans O; KU Leuven, Department of Chemistry, Biochemistry, Molecular and Structural Biology, Celestijnenlaan 200G - Box 2403, Leuven, B-3001, Belgium.
  • Kashyap RS; KU Leuven, Department of Cellular and Molecular Medicine, Laboratory of Protein Phosphorylation and Proteomics, O&N I Herestraat 49 - Box 901, Leuven, B-3000, Belgium; Leuven Cancer Institute (LKI), Laboratory of Protein Phosphorylation and Proteomics, O&N I Herestraat 49 - Box 901, Leuven,
  • Gilles P; KU Leuven, Department of Chemistry, Molecular Design and Synthesis, Celestijnenlaan 200F - Box 2404, Leuven, B-3001, Belgium.
  • De Borggraeve WM; KU Leuven, Department of Chemistry, Molecular Design and Synthesis, Celestijnenlaan 200F - Box 2404, Leuven, B-3001, Belgium.
  • Voet A; KU Leuven, Department of Chemistry, Biochemistry, Molecular and Structural Biology, Celestijnenlaan 200G - Box 2403, Leuven, B-3001, Belgium.
  • Van Lint J; KU Leuven, Department of Cellular and Molecular Medicine, Laboratory of Protein Phosphorylation and Proteomics, O&N I Herestraat 49 - Box 901, Leuven, B-3000, Belgium; Leuven Cancer Institute (LKI), Laboratory of Protein Phosphorylation and Proteomics, O&N I Herestraat 49 - Box 901, Leuven,
Anal Biochem ; 630: 114313, 2021 10 01.
Article em En | MEDLINE | ID: mdl-34302798
Radiometric assays have widely been used for measuring protein kinase activity for decades. In addition, several non-radiometric kinase assay formats have been developed over the years, including luciferase-based and fluorescence-based assays. However, radiometric assays are still considered as the "gold standard" for protein kinase assays, because of their direct readout, high sensitivity, reproducibility, reliability, and very low background signals. These radiometric assays rely on P81 phosphocellulose paper to capture the phosphorylated substrate and wash out unreacted [γ-32P] ATP. However, recently the production of P81 was discontinued by the manufacturer, causing major concern within the protein kinase research community. The advantages of radiometric assays over other kinase assay methods call for an urgent alternative to the discontinued P81 paper. In this report, we demonstrate that the LSA-50 paper is a worthy alternative for radiometric protein kinase assays originally using P81 phosphocellulose paper.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Papel / Proteínas Quinases / Radiometria / Celulose Idioma: En Revista: Anal Biochem Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Bélgica

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Papel / Proteínas Quinases / Radiometria / Celulose Idioma: En Revista: Anal Biochem Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Bélgica