Fig. 6
From: An automated workflow to screen alkene reductases using high-throughput thin layer chromatography
Application of the screen to a codon-saturation mutagenesis library, L377X. a TLC plate for product analysis. The first 48 lanes on the left represent activities in mutants contained in rows A–D of a 96-well plate format; the other 48 lanes on the right represent activities in mutants contained in rows E–H. Protein concentration measured in each well via Bradford assay is denoted under each TLC lane. Following chromatographic separation, NBP-treated plates were heated in an oven at 150 °C for 10 min and developed with triethylamine. b Comparison of Rf values observed in the time-course assay (Fig. 5) denoted as standards with observed Rf values in rows A–D and E–H within the mutant library. While the observed side products Rf (s2) and Rf (s3) migrate slightly differently among plates (dark grey and light grey, respectively), the respective products corresponding to FOH (blue), H2-FOH (green), and H4-FOH (red) at Rf (1), Rf (2), and Rf (3) migrate within error among all TLC plates