Bacterial Cell-Free Protein Expression

• MembraneMax™ Protein Expression System
• Expressway™ Expression System
• Vectors for bacterial cell-free expression
• Antibodies for fusion-tagged proteins
• Ni-NTA and cobalt IMAC purification resins

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).

Key advantages

• High yields—nanogram to milligram   
• Enhanced protein solubility—protein produced in NLP complexes
• Easy purification—His affinity tag
• Complete kit—simply add your gene of interest to get started

Expressway™ Mini and Maxi Cell-Free Expression Systems

The Expressway™ Cell-Free E. coli Expression Systems are designed for in vitro transcription and translation of target DNA to protein in a single reaction. These flexible systems allow production of recombinant protein of interest from an expression construct in as little as 3 hours. Once purified, the resulting recombinant protein is suitable for use in other downstream applications including biochemical, physical, and structural characterization. The system is available in two formats: Expressway™ Maxi (scalable from 20–100 reactions) and Expressway™ Mini (20 reactions). 

The Expressway™ Cell-Free E. coli Expression Systems provide a means to produce high levels of recombinant protein that may be easily detected and purified. These systems offer: 

• High yields—milligram-level protein production within 4–6 hours through the use of a uniquely formulated Feed Buffer
• High-throughput compatibility—reactions can be performed in 96-well plates and once positive expression is detected, scale-up production can be performed
• Optimized TOPO® TA vectors—designed to minimize additional amino acid sequences that may interfere with native protein folding and functionality
• Producing proteins that are toxic to cells

See Expressway™ Mini Cell-Free Expression System products

Expressway™ Lumio™ Expression and Detection System

The Expressway™ Lumio™ Expression and Detection System takes advantage of the Lumio™ recognition sequence, a small, six amino acid sequence (Cys-Cys-Pro-Gly-Cys-Cys). The Lumio™ detection reagent binds the recognition sequence with high specificity and affinity, resulting in a bright fluorescent signal for real-time protein production analysis and immediate in-gel protein detection. In addition, Expressway’s specialized E. coli lysate, derived from a slyD mutant, eliminates nonspecific binding of the Lumio™ Green Detection Reagent to the endogenous SlyD protein and provides optimal background for detection of recombinant proteins. The Lumio™ Green Detection Kit is included in the system for real-time detection of protein synthesis as well as easy in-gel protein detection. 

See Expressway™ Lumio™ Expression products

Find vectors compatible with Expressway™ Cell-Free Expression System and MembraneMax™ Protein Expression Kits.

Choose antibodies for your expressed protein based on the vector for the cell-free protein expression system you're using.

Find immobilized metal affinity chromatography (IMAC) media for the purification of poly histidine-tagged proteins.