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Application notes
Separation of G protein-coupled receptors using ultracentrifuge
G protein-coupled receptors (GPCRs) are located in a cell membrane mainly, and receive many various information such as light, smell, neurotransmitters, hormones and other external signals, and the signals are transduced into the cell via G protein. It is well-known that GPCRs and related proteins are closely linked to various kinds of diseases. They play one of the key roles as research targets for developing new drugs. They have also attracted considerable attention in association with Nobel Prizes twice (The Nobel Prize in Physiology or Medicine 1994 and The Nobel Prize in Chemistry 2012) - The GPCRs are one of the proteins attracting the most global attention today.
Here, we take an example involving the separation of G proteins from lipid raft fractions. Since the lipid raft resists surfactants we dissolved the lipid raft in a surfactant and separated it by isopycnic centrifugation using sucrose density gradient solution. The protein was obtained as a fraction unlike other membrane fractions using different degrees of density Of the liquid.
Fractionation Procedure
CP-WX series ultracentrifuge
P40ST swinging bucket rotor
DGF-U density gradient fractionator
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details.
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