Synthetic Nanodisc Screening Kit MAXI (23x2x50 mg)

Order number: 18295

€1,098.00*

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Description

The versatility of membrane proteins and their composition mirrors the intricate nature of the cell membrane. As a result, it can be a challenge to predict with absolute certainty the optimal copolymer for the solubilization and subsequent stabilization of your membrane protein of interest. Therefore, we have combined our most prominent SMALP, DIBMA, AASTY, and Amphipol products into this practical synthetic copolymer nanodisc Screening Set MAXI to help you find the correct product.
All 23 products are buffered in HEPES to provide maximal comparability between the individual screening assays. In the product description, you can find the complete list of copolymers included in this kit and their specific features.
For general features applicable to all copolymers see Table 1. For more specific features of the individual copolymers see Tables 2 – 6.

Update May 2024: Access the protocol based on the Masterclass by MIT‘s Greg Dodge (PhD): Copolymer Purification Protocol for Screening Approach.

Datasheets

Table 1: Shared Features

This table lists the features that every of the 23 Copolymers share.

Feature
Use	Solubilization & stabilization of membrane proteins
Buffer	HEPES
pH (upon solving in dH₂O)	7.5
State upon delivery	Lyophilized powder
Recommended concentration for use	1% - 5%
Shipping Temperature	Room temperature
Long-term Storage (lyophilized copolymer)	-20°C for several years
Short-term Storage (dissolved copolymer)	2°C - 8°C for several days

Table 2: Included SMALP Products

Feature	SMALP 140	SMALP 140-I	SMALP 200	SMALP 300	Sulfo-SMA
Molecular weight	5 kDa	5 kDa	6.5 kDa	10 kDa	8 kDa
Polymerization Type	Free-Radical	Free-Radical	Free-Radical	Free-Radical	RAFT
Nanodisc Diameter	Diverse	Diverse	Diverse	Diverse	Uniform
Divalent cationic tolerance	< 5 mM	< 100 mM	< 5 mM	< 5 mM	> 100 mM
Absorption at 280nm	Yes	Yes	Yes	Yes	Yes
Solulbility in dH₂O	45%	38%	40%	35%	> 10 %

Table 3: Included SMALP BZ Products

Table 3: Feature	SMALP BZ25	SMALP BZ30	SMALP BZ35	SMALP BZ40
Molecular weight	5.5 kDa	5.5 kDa	5.5 kDa	5.5 kDa
Polymerization Type	RAFT	RAFT	RAFT	RAFT
Nanodisc Diameter	Uniform	Uniform	Uniform	Uniform
Divalent cationic tolerance	< 5 mM	< 5 mM	< 5 mM	< 5 mM
Absorption at 280nm	Yes	Yes	Yes	Yes
Solulbility in dH₂O	> 10 %	> 10 %	> 10 %	> 10 %

Table 4: Included DIBMA Products

Feature	DIBMA 10	DIBMA 12	DIBMA Glycerol	DIBMA Glucosamine	Glyco-DIBMA	Sulfo-DIBMA
Molecular weight	10 kDa	12 kDa	10 kDa	10 kDa	10 kDa	15.7 kDa
Polymerization Type	Free-Radical	Free-Radical	Free-Radical	Free-Radical	Free-Radical	RAFT
Nanodisc Diameter	Diverse	Diverse	Diverse	Diverse	Diverse	Uniform
Divalent cationic tolerance	< 5 mM	< 5 mM	< 50 mM	< 50 mM	< 4 mM	> 100 mM
Absorption at 280nm	No	No	No	No	No	No
Solulbility in dH₂O	> 10 %	> 10 %	> 10 %	> 10 %	> 10 %	> 10 %

Table 5: Included AASTY Products

Feature	AASTY 6-45	AASTY 6-50	AASTY 6-55	AASTY 11-45	AASTY 11-50	AASTY 11-55
Molecular weight	5-6 kDa	5-6 kDa	5-6 kDa	10-11 kDa	10-11 kDa	10-11 kDa
Polymerization Type	RAFT	RAFT	RAFT	RAFT	RAFT	RAFT
Nanodisc Diameter	Uniform	Uniform	Uniform	Uniform	Uniform	Uniform
Divalent cationic tolerance	< 6 mM	< 6 mM	< 6 mM	< 6 mM	< 6 mM	< 6 mM
Absorption at 280nm	Yes	Yes	Yes	Yes	Yes	Yes
Solulbility in dH₂O	> 10 %	> 10 %	> 10 %	> 10 %	> 10 %	> 10 %

Table 6: Included Amphipol Products

Feature	Ultrasolute™ Amphipol 17	Ultrasolute™ Amphipol 18
Molecular weight	7.3 kDa	7.3 kDa
Polymerization Type	Free-Radical	Free-Radical
Nanodisc Diameter	Diverse	Diverse
Divalent cationic tolerance	< 10 mM	< 25 mM
Absorption at 280nm	No	No
Solulbility in dH₂O	> 10 %	> 10 %

Lab Results

SMALP Screening results as a SDS-PAGE — **Figure 1:** Example of a copolymer screening assay. Almost all copolymers included in the Screening Kit Maxi were used here. The screen also includes a few detergents like DDM. Highlighted in red is the protein of interest that should be stabilized and purified. This screening clearly shows that when working with a new protein, an appropriate screening assay is required. High solubilization efficiency does not automatically mean high purification efficiency. Any combination should be screened at best.

BZ SMA Westernblot — **Figure 2:** Shown is the blot activity of a Rho1d4-Tag purified human membrane protein expressed in HEK cells containing 9 transmembrane helices. The membrane protein was solubilized using SMALP BZ25 – 40 as well SMA 200 as control. The chemically modified SMALP BZ series exhibit a superior solubilization and isolation efficiency compared to the compared Copolymers.

Sulfo Dibma Westernblot — **Figure 3:** Shown is the blot activity of a Rho1d4-Tag purified human membrane protein expressed in HEK cells containing 9 transmembrane helices. The membrane protein was solubilized using Sulfo DIBMA as well as other copolymers from the DIBMA family. Besides the efficient solubilization and purification of the membrane protein, the copolymer belt of the resulting nanodiscs is electroneutral.

Sulfo Polymers Net Charge — **Figure 5:** ζ-potential of DMPC nanodiscs formed with different copolymers. Ultrasolute 18, DIBMA 10 and SMA 200 nanodiscs reveal a strong negative net charge whereas nanodiscs formed with Sulfo-DIBMA and Sulfo-SMA reveal a nearly electroneutral nanodisc compared to non-sulfonated controls. ζ-potential was measured in 150 mM NaCl, 20 mM HEPES at pH 7.5 and a Polymer/DMPC weigth ratio of 3/1.

Video

Watch our video on the use of Copolymers for membrane protein solubilization. It demonstrates the use with SMALP.

FAQ

Is DIBMA from Cube Biotech ready to use?

Yes, our DIBMA is ready to use. You can start directly with solubilization. Read our protocol for more information.

Which pH is suitable for DIBMA?

For DIBMA a pH of 7.5 is recommended. DIBMA does not solubilize if the pH is smaller than 6.5.

Which concentrations of DIBMA should I use for my protein?

In general, we advise you to add 2,5 % DIBMA to your solution. But the optimal conditions have to be screened by yourself.

I used DIBMA for protein solubilization and a white precipitate appeared - what happened?

Your DIBMA precipitated. You should check your pH and ensure that your pH never drops down to 6.5.

What SMA is the best one to use?

This is a tricky question, as it depends on the membrane protein which SMA performs best, however, SMA 200 is in general the most recommended. Keep in mind that this is no guarantee for it to be the best but in our experience, it is a good allrounder.

What are the recommended concentrations of SMA?

Mix the supplied solutions at a concentration of 1-5% SMALP to the total solution. These conditions can be used as indications. The optimal SMA concentration must be determined separately for each experiment.

How is SMA stored?

Store in cool and dry places, protected from direct sunlight. Long-term storage is recommended at 4°C

The viscosity of my SMA has decreased, what happene

This can occur by storing SMA at low temperatures (as recommended!). Simply warm the SMA solution up to room temperature to restore its original viscosity.

How do I quantify the amount of protein after SMA treatment?

In contrast to DIBMA, SMA absorbs light at 280 nm similar to proteins. Therefore ultraviolet absorption is no option here. However other protein quantification assays are viable options like:

Bicinchoninic Acid (BCA)
Bradford
Folin-Lowry in some cases

Disclaimer

The use of this styrene maleic acid copolymer (SMA) product for the manufacture of styrene maleic acid copolymer - lipid particles (SMALP), and the use of SMALP, are covered by one or more of the following patents owned by Malvern Cosmeceutics Limited: US 8623414, EP 1890675, GB 2426703, AU 2006253886, JP 5142898, IN 261468, CN ZL200680018957.2 and CA 2,611,144 and by the University of Birmingham: US 8754168.

The purchaser is licensed under those patents to use the SMA for the manufacture of SMALP and to use the SMALP for the purpose of research and development of proteins, including their production (including purification and solubilisation), screening testing, analysis, characterisation (including structural analysis and characterisation), including for the purpose of drug screening and for the purpose of in vitro diagnostics, but not for the purpose of delivery of agents to humans or other animals for therapeutic, diagnostic (including in vivo diagnostics) or prophylactic purposes, which uses are specifically prohibited.

Patent Pending
The purchaser is licensed under those patents to use the SMALP BZ; for the manufacture of lipid particles and to use SMALP BZ so manufactured for the purpose of research and development of proteins, including their production (including purification and solubilization), screening, testing, analysis, characterization (including structural analysis and characterization), including for the purpose of drug screening, but not for the purpose of delivery of agents to humans or other animals for therapeutic, diagnostic, prophylactic purposes, which uses are specifically prohibited.