Reconstitution of hTRPV6 protein into amphipols
This protocol is extracted from research article:
Mechanism of calmodulin inactivation of the calcium-selective TRP channel TRPV6
Sci Adv, Aug 15, 2018; DOI: 10.1126/sciadv.aau6088

hTRPV6 was incorporated into amphipols as described previously (29). Briefly, for reconstitution in A8-35 amphipols (Anatrace #A835), purified hTRPV6 in the presence of 20 μM CaM and 5 mM Ca2+ was mixed with amphipols in a 1:3 mass ratio (protein/amphipols) and incubated for 3 hours with rotation at 4°C. Subsequently, 7 to 8 mg (per 0.5 ml of protein/amphipol mixture) of Bio-beads SM2 (Bio-Rad) prewet in buffer containing 20 mM tris (pH 8.0), 150 mM NaCl, and 1 mM β-mercaptoethanol was added to the protein/amphipol mixture to facilitate the reconstitution of TRPV6 into amphipols. The mixture was rotated for ~20 hours at 4°C, and the amphipol-solubilized TRPV6 was purified as described above.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.