Expressway™ Mini Cell-Free Expression System
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Invitrogen™

Expressway™ Mini Cell-Free Expression System

The Expressway™ Mini Cell-Free Expression System utilizes an efficient coupled transcription and translation reaction to produce active recombinant protein. ThisRead more
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Catalog number K990100
Price (USD)
850.00
Each
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Price (USD)
850.00
Each
Add to cart
The Expressway™ Mini Cell-Free Expression System utilizes an efficient coupled transcription and translation reaction to produce active recombinant protein. This system eliminates the time-consuming steps of cell-based protein production including transformation, cell culture, and expression optimization. In just two hours, you can produce protein suitable for an array of downstream applications and functional studies. The Expressway™ Mini Cell-Free Expression System offers flexibility in experimental design, allowing you to
• Synthesize proteins from both circular plasmid and linear templates (e.g. PCR products)
• Perform high-throughput synthesis of proteins
• Express genes toxic to in vivo systems

How it works:
The Expressway™ Mini Cell-Free Expression System provides all the components you need for optimal cell-free protein production. The kit includes an E. coli extract containing the cellular machinery required to drive transcription and translation. An IVPS (in vitro protein synthesis) reaction buffer and an amino acid module are also included in the kit to provide the required amino acids and an ATP-regenerating system. Simply mix the reaction buffer, amino acid mix, methionine (labeled or unlabeled), T7 Enzyme Mix, and your DNA template (containing a T7 promoter) with the E. coli extract. As the DNA template is transcribed, the 5’ end of the mRNA is bound by ribosomes and undergoes translation as the 3’ end of the template is still being transcribed (Figure 1).


Functional protein made easy:

The Expressway™ Mini Cell-Free Expression System has been optimized to produce high yields of full-length, functional protein from E. coli cell lysates (Figure 2). A proprietary feed buffer formulation is provided with the system for increased protein yields.


Contents and Storage:
The Expressway™ Mini Cell-Free Expression System is designed to perform twenty 50-μl reactions or one 1-ml reaction. This system includes the IVPS E.coli Extract, IVPS reaction buffer, feed buffer, 19 amino acid mix, methionine, DNase/RNase-free water, RNase A, T7 Enzyme Mix, 2-ml reaction tubes, and a positive expression control vector. Suggested vectors for use with the Expressway™ Mini Cell-Free Expression System are listed below. Store vectors at -20 °C. Store other kit components at -80 °C.
WARNING: Reproductive Harm - www.P65Warnings.ca.gov
For Research Use Only. Not for use in diagnostic procedures.
Specifications
PromoterT7
Product TypeCell-Free Expression System
Expressed Protein TypeWater Soluble Protein
Expression SystemCell-Free
Expression MechanismCell-Free Expression
Quantity1 kit
Product LineExpressway™
Unit SizeEach

Frequently asked questions (FAQs)

I accidentally stored my E. coli slyD-Extract, E. coli Reaction Buffer (-A.A.), and 2X feed buffer at room temperature. Can I still use them?

Unfortunately, this may result in a loss of activity.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

I've run out of the T7 RNA polymerase for my cell-free expression. What do you suggest I use?

We would recommend using T7 RNA polymerase (Cat. No. 18033019, 50 U/µL). Use 1-1.5 µL in a 50 µL reaction system.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

I'm getting smearing after running my cell-free expression reaction on a gel.. What could be the cause of this?

Smearing may occur if samples for the following reasons:

- Samples were not precipitated with acetone: precipitate proteins with acetone to remove background smearing.
- Too much protein was loaded: reduce the amount used.
- The gel itself was not clean: rinse the gel briefly before exposing to film.
- Ethanol was present in the protein synthesis reaction: make sure that any residual ethanol is removed during DNA purification.
- Check the date of your pre-cast gels: do not use gels after the expiration date.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

I'm seeing a ladder of small-sized products after running my reaction on a gel when using the Expressway system. Why is this?

There may be several reasons for why this is occurring. The most common are: proteolysis, degradation of DNA and/or RNA templates (truncated templates will generate truncated protein products), internal initiation (if there are many methionines and internal RBS-like sequences in the gene, the ribosome may initiate translation from the wrong methionine), premature termination, translational pausing, frequent rare codon usage, complicated secondary structure of RNA, and others. This can also happen if proteins are denatured for too long, or not enough SDS was added to the 1X SDS-PAGE sample buffer.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

With a cell-free expression system, I'm getting good protein yield, but it has low biological activity. What can I do?

- Your protein may not be folding properly: try to reduce the incubation temperature to as low as 25 degrees C during synthesis.
- You may require post-translational modification of your protein: the Expressway system will not introduce post-translational modifications to the recombinant protein.
- Your synthetic protein may require co-factors for complete activity: try adding required co-factors to the protein synthesis reaction.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.