MembraneMax™ Protein Expression Kits
Membrane proteins play critical roles in cell-to-cell contact, surface recognition, cytoskeleton contact, signaling, enzymatic activity, and transport. Because of their varied cellular functions, they are ideal drug targets. However, membrane proteins are challenging because their expression can be toxic to cells or cause inclusion bodies, which limit protein yield and tedious optimization makes purification difficult and time consuming.
MembraneMax™ Protein Expression Kits overcome these challenges, allowing you to produce high yields of soluble (dispersed) membrane proteins using the MembraneMax™ reagent—a planar phospholipid membrane bilayer surrounded by a scaffold protein (also called a nanolipoprotein particle or NLP).
- High yield--nanogram to milligram
- Soluble membrane protein–NLP complexes
- Simple purification with His tag
- Complete kit--simply add your gene of interest to get started
Figure 1: Soluble membrane protein expression achieved using MembraneMax™ HN Kit
Figure 2: Overview of same day protocol
Figure 3: Expression and activity data for Bacteriorhodopsin protein using MembraneMax™ Kit
Figure 4: Data showing insertion of Membrane protein into MembraneMax™ Reagent
Figure 5: Activity data for the EMRE membrane protein expressed using MembraneMax™ Kit
High yield for your downstream analysis
Expression of membrane protein is often very difficult due to insolubility in bacterial systems, in vivo toxicity or simply low yield in mammalian systems. Using our Ultimate™ ORF collection as a source for membrane proteins, we tested membrane proteins representing differing sizes (8 kDa to 51kDa) and complexity (2 to 7 transmembrane domains). With the addition of the MembraneMax™ reagent, the majority of proteins were expressed >0.1mg/ml of protein (Table 1), which is sufficient and suitable for nearly every downstream application (Table 2).
Table 1—High yields achieved using the MembraneMax™ system.
|Membrane Protein (GenBank Accession Number)||Yield (mg/ml)|
|Glycophorin E (NM_002102.2)||0.30|
|b2 adrenergic receptor (NM_000024.3)||0.30|
Shown above is a small subset of proteins that we have expressed using the MembraneMax™
Protein Expression Kit. Over 64% of the proteins expressed at >0.1 mg/ml of reaction (n = 32).
Table 2—Downstream applications for analysis of membrane proteins.
|Ligand binding or functional assays|
|Nuclear magnetic resonance (NMR)|
|Protein array construction|
Enhanced protein solubility
One of the most pervasive problems in protein expression is the insolubility of recombinant protein, often seen as inclusion bodies. This is especially acute when working with membrane proteins, which have both hydrophilic and hydrophobic domains as well as complex folding motifs. MembraneMax™ Protein Expression Kits use the novel MembraneMax™ reagent to enhance the solubility of overexpressed membrane proteins ranging in size and complexity (Figure 1).
Scalable and easy to use
Simply combine your T7 E. coli expression vector encoding your membrane protein with Expressway™ cell-free E. coli extract and MembraneMax™ reagent, incubate for 2 hours and purify (Figure 2). Within a day, you can generate milligrams of soluble membrane protein for downstream analysis, without detergents or tedious reconstitution protocols. MembraneMax™ Protein Expression Kits contain all the necessary reagents for successful expression of your membrane protein; the cell-free format allows high-throughput expression screening of multiple samples, or large-scale production of a single membrane protein.
Simple purification method
Purification challenges such as tedious and time-consuming testing of various detergents and extraction conditions to remove other endogenous membrane proteins while maintaining your protein's stability and function have notoriously impeded research on membrane proteins. In contrast, MembraneMax™ HN kits contain a His-tagged MembraneMax™ reagent, which simplifies purification without having to add a tag to your membrane protein. You also have the flexibility to design your DNA template with your favorite purification tag at the N- or C-terminus of your membrane protein, which when used with the MembraneMax™ HN module enables tandem purification for very clean membrane protein–NLP complexes. Whether you choose to work with a native membrane protein or a tagged version, our MembraneMax™ kits give you the flexibility you need
Take advantage of our special introductory pricing--click on the catalog number to see your price!
|Kit||Features||20 rxns||100 rxns|
|MembraneMax™ Protein Expression Kit||
|MembraneMax™ HN Protein Expression Kit||
Find your membrane protein from our Ultimate™ ORF Collection