I am currently interested in identifying some receptor proteins on the plasma membrane of Sf9 cells. However, I find it difficult getting hold of any protocol that I can use to extract these proteins while preserving them in their native form. Any suggestions please?
The first step could be to purify the plasma membranes using preparative sucrose gradient ultracentrifugation of disrupted cells (sonication can be used). The plasma membranes are the lightest membrane fraction. The membrane fraction can be stored frozen until the extraction.
A non-ionic or zwitterionic detergent should be used to extract the membrane proteins. Commonly used detergents for this purpose are n-octyl-beta-D-glucopyranoside (octylglucoside), and dodecylmaltoside. I have also used sulfobetaine detergents such as Zwittergent 3-12 and 3-14. You can experiment with different concentrations of detergent to find what works best.
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