Infection of HEK293 Mammalian Cells with P2 Baculovirus for Large-scale Protein Expression
2:42
Purification of hTRPC3 Protein from the Frozen Cell Pellet
5:26
Results: Structural Determination of hTRPC3
7:36
Conclusion
Transcription
This protocol enabled us to solve the structure of human TRPC3, a member of an important, but understudied family of ion channels. The main advantage of this technique is the high efficiency with which it can be applied to express and purify a var
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This protocol describes techniques used to determine ion channel structures by cryo-electron microscopy, including a baculovirus system used to efficiently express genes in mammalian cells with minimum effort and toxicity, protein extraction, purification, and quality checking, sample grid preparation and screening, as well as data collection and processing.