Synthetic Nanodisc Screening Kit MINI, (8x2x50 mg)

Order number: 18290

€442.00*

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Description

Finding the best-performing synthetic copolymer nanodisc for your respective nanodisc can be tricky. Especially if you have never worked with your current membrane protein of interest before. For these cases, we have compiled our most prominent SMALP and DIBMA products into this practical Synthetic Nanodisc Screening Kit for your convenience!

All 8 products are buffered in HEPES to provide maximal comparability between the individual screening assays. In the product description, you can find the list of copolymers included in this kit. DIBMA Glucosamine, Sulfo-DIBMA, and Sulfo-SMA provide the highest cation tolerance of all.

Figure 1 shows the necessity for a screening before choosing a SMALP product. Note that the detergent LMNG may provide the highest protein yield, but it must be added to the purified protein constantly and it might interfere with the SPIKE protein's function.

If the MINI Kit's contents are not enough, we suggest you to try the MAXI Screening Kit instead.

Datasheets

Small Scale Screening Protocol for Copolymers Protocol

Membrane Protein Solubilization Protocol with Copolymers & Detergents

Table 1: Shared Features

This table lists the features that every of the 8 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
Storage conditions (short-term, dissolved)	2°C - 8°C for several days
Storage conditions (long-term, lyophilized)	-20°C for several years

Table 2: Included SMALP Products

Feature	SMALP 140	SMALP 200	SMALP 300	Sulfo-SMA
Molecular weight	5 kDa	6.5 kDa	10 kDa	9 kDa
Polymerization Type	Free-Radical	Free-Radical	Free-Radical	RAFT
Nanodisc Diameter	Diverse	Diverse	Diverse	Uniform
Divalent cationic tolerance	< 5 mM	< 5 mM	< 5 mM	> 100 mM
Absorption at 280nm	Yes	Yes	Yes	Yes
Solulbility in dH₂O	45%	40%	35%	> 10 %

Table 3: Included DIBMA Products

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

Citations

Solubilized Protein	Year	Author
Several different proteins	2021	Overduin M., Trieber C., Prosser R.S., Picard L-P., Sheff J.G.
Serveral different membrane proteins	2022	Glueck D., Grethen A., Das M., Ogochukwu M. P., Patallo E. P., Meister A., Rajender R., Kins S., Räschle M., Victor J., Chi C., Etzkorn M., Köck Z., Bernhard F., Babalola J.O., Vargas C., Keller S.
ATP-binding cassette (ABC) transporter	2023	Dimitrova V.A., Song S., Karagiaridi A., Marand A., Pinkett H.W.
Env-protein of HIV	2023	Zhou R., Zhang S., Nguyen H.T., Ding H., Gaffney A., Kappes J.C., Smith III. A.B.S, Sodroski J.G.

Lab Results

SMALP Screening results as a SDS-PAGE — **Figure 1:** Example of a SMALP screening assay. Different SMALPs compared in after solubilisation & purification of Rho1-tagged SPIKE protein of coronavirus.

Sulfo Dibma Westernblot — **Figure 2:** Shown is the blot activity of a Rho-Tag purified human membrane protein expressed in HEK cells containing 9 transmembrane helices. The membrane protein was solubilized using Sulfo-SMA as well as other copolymers from the SMA family. The electroneutral Sulfo-SMA exhibits the highest solubilization and purification efficiency of all testes SMAs. The purified membrane protein is stabilized in an electroneutral copolymer belt.

Sulfo Polymers Net Charge — **Figure 4:** 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. But it can also be applied to DIBMA as both polymers share the same protocol.

FAQ

DIBMA

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.

SMA

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.

The use of SMALP 502-E within the United States is not covered by our current license and this product can therefore not be made available for US purchasers.