Fig. 1.

CD81 can be solubilised using SMA polymers or conventional detergent. Membrane preparations from CD81-expressing P. pastoris cells (30 mg/ml wet pellet weight) were solubilised with 2% (w/v) DDM, 2.5% (w/v) SMA polymer or PBS (negative control) for 1 h at room temperature. A) Image showing the change in appearance as SMA clarifies the membrane suspension compared to PBS control. B) Optical density measurements of the samples measured at 600 nm. Data are mean ± SD, n = 3. C) Following solubilisation, samples were centrifuged at 100,000g for 20 min at 4 °C. Solubilised protein in the supernatant (S) was harvested. Insoluble material in the pellet (P) was resuspended in an equal volume of purification buffer supplemented with 2% (w/v) SDS. Samples were analysed by Western blot using an anti-CD81 primary antibody (mAb 2s131). D) Average CD81 solubilisation efficiency. Data are mean ± SD, n = 3.