Workflow from Scientific Research

General TPP workflow. Cells were treated either with vehicle control or Ciprofloxacin (70 M) in duplicates for 1 h and cell aliquots were subjected to a temperature gradient with 10 increments. After lysis and ultra-centrifugation to remove aggregated proteins, the soluble fractions were digested by trypsin and the resulting peptides isotopically labelled with TMT 10-plex with one distinct channel per temperature. After combination of all 10 TMT channels per sample and offline HILIC fractionation, the samples were measured by LCMS/MS and the raw data analyzed to infer protein melting curves.

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