Synthetic Nanodisc Screening Kit MAXI (23x2x50 mg)

Order number: 18295

€1,098.00*

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Description

Membrane Protein with AASTY nanodisc
Membrane Protein with Amphipol nanodisc
Membrane Protein with DIBMA nanodisc
Membrane Protein with SMALP nanodisc
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 synthetic polymer 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 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 polymers included in this kit and their specific features.
For general features applicable to all polymers see Table 1. For more specific features of the individual polymers see Tables 2 – 6.

Table 1: Shared Features

This table lists the features that every of the 23 polymers 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 polymers.
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 polymers from the DIBMA family. Besides the efficient solubilization and purification of the membrane protein, the polymer belt of the resulting nanodiscs is electroneutral.
SMALP Screening results as a SDS-PAGE
Figure 4: Example of a SMALP screening assay. Different SMALPs compared in after solubilisation & purification of Rho1D4-tagged SPIKE protein of coronavirus.
Sulfo Polymers Net Charge
Figure 5: ζ-potential of DMPC nanodiscs formed with different polymers. 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 synthetic polymers 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.