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. Author manuscript; available in PMC: 2021 Dec 1.
Published in final edited form as: Anal Biochem. 2020 Aug 4;610:113887. doi: 10.1016/j.ab.2020.113887

Figure 1. Improved purification and biophysical characterization of the Torpedo californica-nicotinic acetylcholine receptor in lipid-like detergent complexes (Tc-nAChR-DC).

Figure 1.

The scheme presents the step by step the procedure used for the preparation of a highly pure, stable and functional nAChR-LFC-16 complex. The most relevant innovation involves a sequential purification system, which consists of (5 & 7) two steps of FPLC-affinity purification and (8) one step of FPLC-size exclusion chromatography. (9) Using SDS gels and Microfluidic Capillary Gel Electrophoresis (MCGE) Bioanalyzer, for the evaluation of protein purity. (10) We performed circular dichroism (CD) for the secondary structure predictions and (11) Fluorescence Recovery After Photobleaching (FRAP) using confocal microscopy for stability measurements in LCP. (12) Finally, we used the TEVC to access the functionality of the purified nAChR-LFC-16 injected into Xenopus Laevis oocyte.