PRODUCT

E. coli Cell-Free Protein Expression Kit

Fast cell-free protein expression from circular & linear DNA

Go from genes to proteins in hours. This fast cell‑free protein synthesis system uses optimized E. coli lysates and supports both circular and linear DNA templates to produces up to 20 µg of functional protein in ~4 hours. Ideal for rapid screening, parallel expression, and workflow scaling in tubes or microplates.

High yield E. coli lysate drives efficient coupled transcription/translation, generating up to 20-µg protein per 50-µL reaction in ~4 h
Supports linear and plasmid DNA for rapid construct screening without cloning, improving early expression workflows
Flexible format works in microplates or tubes, enabling scalable, parallelized protein production
Suitable for optimization studies, including chaperone or additive screening to enhance active protein yield

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E. coli Cell-Free Protein Expression Kit (6 Reactions)

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Specifications Description Quality FAQ Resources

Product Number	CF31ECXHY
Delivery Format	Tubes
Source	E. coli
Shipping Conditions	Dry Ice
Storage Temperature	-20 C
Stability	24 months from production; refer to expiration dates for details
Function	Coupled in vitro transcription and translation; Cell-free protein expression/synthesis; High-efficiency protein production from both linear and circular templates.
For Use In	Microgram-scale protein expression; Rapid gene contruct screening without cloning; Parallel protein expression; Functional protein studies
Intended Use	This product is intended for research use only

Express Proteins Faster

The E. coli Cell-Free Protein Expression Kit is an advanced solution for efficient in vitro protein expression, utilizing a high-yield E. coli lysate for coupled transcription and translation. This kit is capable of producing up to 20 μg of protein per 50 μL reaction in as little as 4 hours, making it ideal for rapid protein-expression screening and optimization. Compatible with both linear templates and plasmids, the kit offers flexibility for a variety of experimental setups.

Designed for convenience, it includes all necessary components for protein expression in either microplates or microtubes, supporting both small-scale and high-throughput workflows. The system also allows for optimization of protein production by incorporating additional reagents such as chaperones or detergents, ensuring active and functional protein yields.

Key Features

Rapid Protein Expression: Achieve protein yields of up to 20 μg per reaction in just 1–4 hours.
Template Compatibility: Works seamlessly with both linear templates and plasmids, enabling quick screening without the need for cloning.
High Yield Efficiency: Optimized biochemistry ensures reliable and consistent protein production.
Streamlined & Flexible Setup: Simplified reaction preparation suitable for use in microplates or microtubes, with adaptability for high-throughput applications.

Applications

Membrane and Soluble Protein Expression: Capable of expressing both standard soluble proteins and membrane proteins.
Rapid Protein Screening: Ideal for fast, parallel expression of proteins at the microgram scale.
Functional Studies: Facilitates the assignment of biological functions to genes through efficient protein production.
Optimization of Expression Conditions: Supports the addition of reagents like chaperones or detergents to enhance active protein yields.

Custom Component Options

We offer flexible customization to meet your specific needs:

Volume: Optimize packaging sizes to match your application scale.
Concentration: Customize concentrations for optimal performance.
Formulation: Available in custom formats to suit your protocols.

Quality

In vitro protein expression products are manufactured using high quality chemicals and materials.

Component Testing
Each component is tested to ensure the composition and quality meet stated specifications.

Performance Testing
Each product is tested to ensure it meets the stated performance specification.

FAQ

Answers to Commonly Asked Questions

How much protein can I produce with your kits?

The E. coli Cell-Free Protein Expression Kit is designed for small-scale reactions and can produce up to 20 µg of protein in just 4 hours. The protein expression yield of a given gene is influenced by multiple factors, including and not limited to its mRNA structure, the amount of template DNA utilized per reaction, and the use or not of certain additives. Therefore, expression condition optimization is strongly recommended for those who seek to maximize the yield.

Overall, this kit makes it ideal for rapid screening and optimization of expression constructs, where smaller amounts of protein are sufficient for analysis. The kit delivers high-quality, functionally active proteins that are ready for downstream applications such as enzymatic assays, binding studies, or initial structural analysis.

What templates are compatible with your kits?

The E. coli Cell-Free Protein Expression Kit is compatible with both linear DNA templates and plasmids. Linear DNA templates can be generated quickly using PCR, making them ideal for rapid screening and testing multiple constructs in parallel. Plasmid templates are also supported and can be used for small-scale reactions. Regardless of the template type, it must include essential regulatory elements such as a T7 promoter, a ribosomal binding site (RBS), and a T7 terminator to ensure efficient transcription and translation in the cell-free system.

Do I need special equipment to use these kits?

No special equipment is required to use the E. coli Cell-Free Protein Expression Kit. A standard laboratory incubator that can host 200-µL PCR tubes is sufficient to maintain the reaction at the recommended temperature of 30°C. The simplicity of the setup makes this kit accessible for any lab, even those without specialized equipment. Unlike medium-scale systems, this kit does not require specific shaking or additional devices, making it a convenient choice for small-scale protein expression.

Can I add supplements to the reaction to improve protein quality?

Yes, the E. coli Cell-Free Protein Expression Kit allows the addition of supplements to improve protein quality including solubility, yield, and functionality. For example, detergents like DDM or CHAPS can be added to stabilize membrane proteins, while stabilizers like sorbitol can enhance the solubility of non-membrane proteins. Chaperones such as DnaK or GroE can also be included to assist in proper protein folding. Please refer to Appendix 3 of the User Guide for more details.

The small reaction volume (50 µL) makes this kit an excellent platform for testing the effects of supplements before scaling up to larger reactions.

If you have additional questions please connect with a member of our team.

RESOURCES

Download the User Guide

CF31ECXHY User Guide

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