Characterization of samples purified in a mixture of CHAPS and Fos14 detergents.

P. pastoris membranes were solubilized and purified using a two-step purification approach (anti-flag affinity chromatography followed by SEC). A, Samples were solubilized in a mixture of 0.25% Fos14/0.1% CHAPS and purified in presence of 0.1% Fos14/0.1% CHAPS. 1: SDS-PAGE of SEC elution fractions F17 and F22 colored with Coomassie Blue (left) and revealed by anti-Flag immunodetection (right). 2: Saturation ligand binding experiments with [³H]-melatonin on SEC elution fraction F22. 3: Negative staining electron microscopy on SEC elution fraction F22. B, Samples were solubilized in a mixture of 0.25% Fos14/1% CHAPS, purified in the presence of 0.1% Fos14/0.5% CHAPS, and analyzed by negative staining electron microscopy.