Fig. 3: Detection of CnTagged proteins in cell extract via in-gel fluorescence and Coomassie stain of the same SDS-gels.
From: Specific, sensitive and quantitative protein detection by in-gel fluorescence
a Small quantities (125 fmol) of CnTagged proteins were mixed with large quantities (~20 µg) of E. coli cell extract and visualized via in-gel fluorescence (left) or with Coomassie (right). The shown proteins are Single domain Antibody (SdAb), Ubiquitin (Ub), GroES, Glutathione-S-Transferase (GST), Ubiquitin-conjugating enzyme (E2), Ubiquitin-activating enzyme (E1), Heat-shock protein 20 kDa (Hsp20), Proteasome subunit Alpha (PsmA), Outer membrane phospholipase A1 (OmpLA) and Cyclophilin A (CyP). The molecular weights and native assembly states of the proteins are indicated. The employed fluorophore-Connectase conjugate (N-Cnt, see control in lane 1) is only visible as a very faint band in presence of CnTagged POI, indicating an effective transfer of the fluorophore. In high-percentage gels, residual fluorescent peptide substrate (~3 kDa) can be seen at the bottom of the gel (see Supplementary Information). Source data are provided as a Source Data file. b Small quantities (125 fmol) of CnTagged E1 protein were mixed with large quantities of Spodoptera frugiperda SF9, human HEK293, Bacillus subtilis, Pseudomonas fluorescens or Sulfolobus solfataricus cell extracts and visualized via in-gel fluorescence (left) or with Coomassie (right). Source data are provided as a Source Data file.
