バクテリア無細胞タンパク質発現

ダウンストリーム分析用の高収率

膜タンパク質の発現は多くの場合、バクテリアシステムにおける不溶性、in vivo毒性、または単に哺乳類システムにおける低収率によって、大変難しくなります。弊社のUltimate™ ORFコレクションを膜タンパク質ソースとして使い、異なるサイズ( 8kDa ～51kDa)および複雑さ(2～7 膜貫通領域)を示す膜タンパク質を検査しました。MembraneMax™試薬の追加で、タンパク質の大部分は>0.1mg/mLのタンパク質を発現します（表を参照）。これはほとんど全部のダウンストリーム用途、例えば抗体産生、結晶学、免疫沈降法、リガンド結合または機能アッセイ、質量分析法、NMR、タンパク質アレイ構築などに十分かつ適しています。

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.

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. 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.

See a detailed protocol for MembraneMax™ protein expression

See the MembraneMax™ Protein Expression Kits user guide