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Optimized mass spectrometry-based proteomics workflow for the study of H. volcanii . Cells grown in glucose synthetic media were lysed following three different protocols: methanol-chloroform extraction (MCE), acid guanidinium thiocyanate-phenol-chloroform extraction (AGPC), and SDS-based cell lysis. The most efficient protein extract, based on the protein yield, was digested following the s-Trap protocol, and peptide samples were analyzed in timsTOF instruments using dia-PASEF. Figure was generated using BioRender.com

Journal: microLife

Article Title: Proteome characterization of single gene deletion mutants lacking zinc finger µ-proteins in Haloferax volcanii

doi: 10.1093/femsml/uqag005

Figure Lengend Snippet: Optimized mass spectrometry-based proteomics workflow for the study of H. volcanii . Cells grown in glucose synthetic media were lysed following three different protocols: methanol-chloroform extraction (MCE), acid guanidinium thiocyanate-phenol-chloroform extraction (AGPC), and SDS-based cell lysis. The most efficient protein extract, based on the protein yield, was digested following the s-Trap protocol, and peptide samples were analyzed in timsTOF instruments using dia-PASEF. Figure was generated using BioRender.com

Article Snippet: Eluting peptides were analyzed on a hybrid trapped ion mobility spectrometer (TIMS)—quadrupole time of flight (Q-ToF) mass spectrometer timsTOF Pro II (Bruker Daltonics) coupled to the nano-HPLC by a CaptiveSpray nano-electrospray ion source.

Techniques: Mass Spectrometry, Extraction, Lysis, Data-independent acquisition, Generated