Nanodisc Assembly Kit MSP2N2-His POPC

Order number: 26273

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Phospholipids

DMPC

POPC

Quantity

Description

Membrane scaffold proteins derive from apolipoprotein (apo; A-I). They are used in membrane protein sciences to stabilize membrane proteins in a lipid environment after the removal of the cell membrane. The size of the nanodisc is determined by the MSP that was used to create the disc. MSP2N2 (His-tagged) creates nanodiscs of ~17 nm in diameter.

This product is a MSP nanodisc assembly kit. It contains a vial of lyophilized membrane scaffold protein MSP2N2 (His-tagged) and a vial of DMPC or POPC phopshospholipid to assemble a MSP nanodisc.

With this assembly kit both cellular as well as cell-free expressed proteins can be stabilized in MSP nanodiscs. Just use the corresponding protocol for each purpose (see below). The MSP protein contains a His-tag, keep that in mind when choosing the affinity tag for your membrane protein.

Other MSP products or related sites by Cube Biotech include:

MSP nanodisc schematic overview — **Figure 1:** Schematic of a MSP nanodisc. A Membrane-scaffold-protein (MSP, green) forms a disc-shaped structure with some phospholipids.

Datasheets

Feature - MSP Protein
Origin of MSP Protein	Human
Purity	> 90% (Determined by SDS-PAGE)
Number of amino acids	392
Molecular Mass	45,670 Da
Extinction coefficient (in water) ε₂₈₀	44,920 M^-1cm^-1
Buffer	20 mM TRIS pH 7.4; 100 mM NaCl; 0.5 mM EDTA
Sequence (Human origin), for the other origins please send request	GHHHHHHH DYDIPTT ENLYFQG STFSKLREQLG PVTQEFWDNLEKETEGLRQEMS KDLEEVKAKVQ PYLDDFQKKWQEEMELYRQKVE PLRAELQEGARQKLHELQEKLS PLGEEMRDRARAHVDALRTHLA PYSDELRQRLAARLEALKENGG ARLAEYHAKATEHLSTLSEKAK PALEDLRQGLL PVLESFKVSFLSALEEYTKKLNTQ GT PVTQEFWDNLEKETEGLRQEMS KDLEEVKAKVQ PYLDDFQKKWQEEMELYRQKVE PLRAELQEGARQKLHELQEKLS PLGEEMRDRARAHVDALRTHLA PYSDELRQRLAARLEALKENGG ARLAEYHAKATEHLSTLSEKAK PALEDLRQGLL PVLESFKVSFLSALEEYTKKLNTQ
Affinity tag	N-Terminal His-tag
Shipping Temperature	Room Temperature
Storage of lyophilized protein	-20°C for several months
Storage of reconstituted protein	2-8°C for several days
Helpful literature references	Bayburt, T.H. et al. Reconstitution and imaging of a membrane protein in a nanometer-size phospholipid bilayer. J. Struct. Biol. (1998), 123(1):37-44 Civjan, N.R. et al. Direct solubilization of heterologously expressed membrane proteins by incorporation into nanoscale lipid bilayers. BioTechniques (2003) 35:556-563 Hagn, F. et al. Optimized phospholipid bilayer nanodiscs facilitate high-resolution structure determination of membrane proteins. J.Am.Chem. Soc. (2013), 135:1919-1925 Proverbio D., et al. Functional properties of cell-free expressed human endothelin A and endothelin B receptors in artifical membrane environments. Biochim.Biophys. Acta (2013), 1828(9):2182-92 Roos, C., et al. 2014, High-level Cell-free production of membrane proteins with Nanodiscs. In: Alexandrov, K., and Johnston W.A. (eds) Cell-free protein synthesis: Methods and Protocols. Methods in Molecular Biology, vol. 1118, Springer Science+Business Media.

Feature - Phospholipids
CAS-Number	DMPC: 18194-24-6 POPC: 26853-31-6
Molecular Mass	DMPC: 677.93 Da POPC: 760.08 Da
Chemical Formula	DMPC: C₃₆H₇₂NO₈P POPC: C₄₂H₈₂NO₈P

Lab Results

Watch our video tutorial about the creation of MSP nanodiscs. It explains the whole process, starting with the initial solubilzation using detergents. The protein that was stabilized in this demonstration was Bacteriorhodopsin.

We also recommend our video guide to nanodiscs in general.

FAQ

Can I get the datasheet for the MSP2N2 (His-tagged) nanodisc assembly Kits?

Yes. Just download or request them here.

Is this the correct MSP product for my membrane protein?

This cannot be answered easily as every membrane protein performs differently with each nanodisc size and phospholipid. It usually requires a screening process before. Remember that membrane proteins with fewer transmembrane domains (TMD) require a smaller nanodisc, while a high number of TMDs need a larger nanodisc.

Can I shorten the cell-free stabilization protocol?

In a way. You can use our already pre-assembled MSP nanodiscs to skip the actual time consuming assembly of the nanodisc.

MSP nanodiscs require solubilization with detergents in before. Do you also offer these?

Yes, we do. Just have a look at them here.

I have never worked with MSP nanodiscs before. Can you tell me more about them?

Of course! In fact we created THIS MSP NANODISC GUIDE PAGE, for this purpose. Have a look at it!

Are there other options to stabilize membrane proteins besides MSP nanodiscs?

Yes, there are. We recommend synthetic copolmer nanodiscs greatly! Have a look at THIS GUIDE PAGE to learn more.

Disclaimer

Our products are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Nanodiscs are protected by US Patents 7,691,414; 7,662,410; 7,622,437; 7,592,008; 7,575,763; 7,083,958; 7,048,949