Expressway™Lumio™ Expression and Detection System, without vector
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Invitrogen™

Expressway™Lumio™ Expression and Detection System, without vector

The Expressway™ Lumio™ Expression and Detection System takes advantage of the Lumio™ recognition sequence, a small, six amino acid sequenceRead more
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Catalog number K990060
Price (USD)
994.00
Each
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Price (USD)
994.00
Each
Add to cart
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 (Figure 1). 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. The Lumio™ vectors (Figures 2 and 3) also feature:

• attR sites for efficient recombination with any attL-flanked Gateway™ entry vector
• N-terminal Lumio™ tag (pEXP3-DEST vector) with TEV cleavage site for efficient removal of the Lumio™ sequence following purification
• C-terminal Lumio™ tag (pEXP4-DEST vector) for easy, and immediate in-gel detection
• T7 promoter, ribosome binding site, and T7 terminator optimally spaced for cell-free protein expression
WARNING: Reproductive Harm - www.P65Warnings.ca.gov
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product TypeLumio Expression and Detection System
Detection LocationIn-Gel Detection
Expressed Protein TypeWater Soluble Protein
Expression SystemCell-Free
Expression MechanismCell-Free Expression
Quantity1 kit
Product LineExpressway™, Lumio™
Unit SizeEach
Contents & Storage
The Expressway™ Lumio™ Expression and Detection System includes the E. coli slyD- Extract, 2.5X IVPS E. coli Reaction Buffer, T7 Enzyme Mix, the Feed Buffer, 19 amino acid mix, Methionine, a control plasmid, DNAse/RNAse-free Water, and a Lumio™ Green Detection Kit for 20 real-time reactions or 100 in-gel applications. In addition to these components, the N-term Expressway™ Lumio™ kit also includes 6 μg of the pEXP3-DEST vector and the C-term Expressway™ Lumio™ kit 6 μg of the pEXP4-DEST vector. Store the vectors and the detection reagent at -20°C. Store all other components at -80°C. Guaranteed stable for 6 months when properly stored.

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.

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