Nanodiscs provide an exciting alternative for the stabilization of membrane proteins. Cube Biotech offers everything you need to get started with this novel technology. There are two options to reconstitute membrane proteins into nanodiscs:
For the reconstitution of membrane proteins that are already isolated and solubilized in detergent, we recommend to use membrane scaffold proteins (MSPs) in combination with phospholipids. A variety of membrane scaffold proteins (MSPs) are available that can be assembled to nanodiscs of different sizes to accommodate proteins with varying numbers of transmembrane domains. His-tagged MSPs allow for easy detection of the nanodisc and even immobilization on SPR chips. On the other hand, if the protein of interest itself carries a His-tag, we recommend to use untagged MSP proteins. For use in cell-free expression reactions, empty nanodiscs are recommended. They can be assembled using our Nanodisc assembly kits, or purchased ready to use. Note that these kits only provide small amounts of nanodisc that are suitable for cell-free expression, and not recommended for direct reconstitution of already purified proteins.
Different nanodisc features in detail
There are several different components that a nanodisc is made from. Each with its own destinct role for the functionality of the MSP nanodisc. The following figure gives an overview of every possible composition and modification of MSP nanodiscs that we offer. In addition to the features listed in figure 1, we also offer DIBMA based synthetic Nanodiscs here.
The size of the MSP Nanodisc depends on the used membrane scaffolding protein. Chosing the correct size of the nanodisc is critical if your essay e.g. cryo-EM requires that only one membrane protein sits in each disc.
Immunization assays require knowledge about the origin of the protein that is used. To cover most situations each of our MSPs can be purchased in one of four different backgrounds.
We offer our pre-assembled nanodiscs and MSP protein tagged and untagged. MSP proteins can be pruchased with and without a poly-His tag. While the pre Assembled nanodiscs can be labled with biotin or not at their phospholipids.
Different membrane proteins prefer different phospholipids in their surroundings to work properly. Sometimes screening is necessary to determine which phospholipid is the correct one. Our store includes the most commenly used phospholipids. All products listed here are pre-assembled nanodiscs. They are usually used to stabilize cell-free expressed
All MSPs can be purchased in wt form or with a D73C mutation for further modifications. An example for this modifications might be Biotinylation.