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. 2020 Nov 16;9(11):401. doi: 10.3390/biology9110401

Figure 2.

Figure 2

Example sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and fluorescence size exclusion chromatography (FSEC) results from a small-scale detergent screen for a bacterial transporter (a–c) and bacterial membrane kinase (d–f). A SRT-300 (Sepax Technologies Inc.) high-pressure liquid chromatography column was used for FSEC. This has the advantage of tolerating higher pressures than standard SEC columns, enabling faster flow rates at 4 °C. 96 samples can be processed in 24 h. (a) Purification in 12 different extraction conditions of a monomeric bacterial transporter (35 kDa) using Ni-NTA resin. (b) Purification of the same bacterial transporter using Talon resin (cobalt). (c) FSEC profiles using tryptophan fluorescence (excitation at 290 nm, emmission at 350 nm) for purifications shown in panel b. (d) Purification in 12 extraction conditions of a multimeric bacterial histidine kinase. SDS-PAGE gel imaged using GFP fluorescence and (e) Coomassie staining. (f) FSEC profile using GFP fluorescence (excitation at 490 nm, emission at 510 nm) of purification in panels (d) and (e). Peaks correspond to monomeric and dimeric protein.