How to make MSP nanodiscs: The protocol
Автор: Cube Biotech
Загружено: 2021-05-31
Просмотров: 4444
MSP nanodiscs are great tools to stabilize & solubilize membrane protein outside the cell membrane for protein purification.
The disc structure is held together by a Membrane-Scaffold-Protein (MSP), a variant of the Apolipoprotein-1 that is involved in the transport of cholesterol in the body.
The buffer and the whole presented protocol can be found here:
https://cube-biotech.com/media/ba/77/...
The demonstrated MSP nanodisc assembly was done using this nanodisc assembly kit:
https://cube-biotech.com/products/mem...
0:00 - Intro
0:12 - What does an MSP nanodisc do?
0:33 - Example: Bacteriorhodosin
0:55 - Setup: The materials
1:16 - Calculating the correct ratios of MSP: lipid : protein of interest
3:43 - MSP nanodisc assembly
4:18 - Removal of detergent via bio beads
5:36 - Purification through SEC
6:55 - Concentrate elution fractions
7:29 - Endcard
This protocol explains an MSP nanodisc assembly example using a MSP1D1-deltaH5 nanodisc with DMPC as the used phospholipid. For other MSP nanodisc - Lipid combinations:
You can find them on matching product sites on our homepage. Example:
https://cube-biotech.com/products/mem...
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