- Cleaner System - sample degradation eliminated
- Easy-to-use - protein expression complete in approximately two hours
- Simple Analysis - protein can often be visualized directly on a Coomassie stained gel
- PURExpress Solution A
- PURExpress Solution B (Minus RF123)
- Control (DHFR) template (10 μl)
The following reagents are supplied with this product:
|PURExpress Solution A|
|PURExpress Solution B|
|Control (DHFR) template|
Advantages and Features
- Quickly generate analytical amounts of protein for further characterization
- Confirmation of open reading frames
- Examination of the effects of mutations on ORFs
- Generation of truncated proteins to identify active domains and functional residues
- Introduction of modified, unnatural or labeled amino acids
- Epitope mapping
- Expression of toxic proteins
- Ribosome display
- Translation and/or protein folding studies
- In vitro compartmentalization
Properties and Usage
Legal and Disclaimers
LicensesPURExpress® is based on the PURE System Technology originally developed by Dr. Takuya Ueda at the University of Tokyo and commercialized as the PURESYSTEM® by BioComber (Tokyo, Japan).
Licensed from BioComber (Tokyo, Japan) under Patent Nos. 7,118,883; WO2005-105994 and JP2006-340694. For research use only. Commercial use of PURExpress® Kit requires a license from New England Biolabs, Inc.
Material Safety Datasheets
- The DHFR control template is now supplied at 125 ng/µl. Use 2 µl for the positive control reaction. Template DNA, particularly plasmid DNA prepared by mini-prep (e.g. Qiagen) is often the major source of RNase contamination. We strongly recommend adding 20 units Murine RNase Inhibitor (NEB #M0314) to each reaction.
- PURExpress DHFR Control Template sequence files: Fasta GenBank
- Storage: All kit components should be stored at -80°C.
- Add Solution B to Solution A, do not dilute Solution B unbuffered. We recommend a starting amount of 250 ng template DNA per 25 μl reaction. The optimal amount of input DNA can be determined by setting up multiple reactions and titrating the amount of template DNA added to the reaction. Typically, the optimal amount will fall in a range of 25–1000 ng template per 25 μl reaction.
- Asahara, H. and Chong, S. (2010). In vitro genetic reconstruction of bacterial transcription initiation by coupled synthesis and detection of RNA polymerase holoenzyme. Nuc. Acid. Res.
- Noto, T., Kurth, H., Kataoka, K., Aronica, L., DeSouza, L., Siu, K., Pearlman, R., Gorovsky, M. and Mochizuki, K. (2010). The tetrahymena argonaute-binding protein Giw1p directs a mature argonaute-siRNA complex to the nucleus. Cell. 140, 692-703.
- Tanner, D., Cariello, D., Woolstenhulme, C., Broadbent, M. and Buskirk, A. (2009). Genetic identification of nascent peptides that induce ribosome stalling. J. Biol. Chem. 284, 34809-34818.
- Talabot-Ayer, D., Lamacchia, C., Gabay, C., and Palmer, G. (2009). Interleukin-33 is biologically active independently of Caspase-1 cleavage. J. Biol. Chem. 284, 19420-19426.
- Feng, Y. and Cronan, J. E. (2009). A new member of the Eschericia coli fad regulon: transcriptional regulation of fadM (ybaW). J. Bacteriol. 191, 6320-6328.
- Solaroli, N., Panayiotou, C., Johansson, M., and Karlsson, A. (2009). Identification of two active functional domains of human adenylate kinase 5. FEBS Lett. 283, 2872-2876.
- When using PURExpress, I was unable to synthesize the control protein?
- When using PURExpress, I was able to synthesize the control protein, but the target sample is not present or present in low yield?
- When using PURExpress, I was able to synthesize the target protein, but full-length product is not major species?
- Detailed FAQs for PURExpress?
- Protein Synthesis Reaction using PURExpress (E6800)
- Analysis of Synthesized Protein using PURExpress (E6800)
- Determination of Protein Synthesis Yield with PURExpress (E6800)
- Purification of Synthesized Protein using Reverse His-tag Purification
- Measurement of 35S-Methionine Incorporation by TCA Precipitation and Yield Determination using PURExpress
- Use of the PURExpress In Vitro Protein Synthesis Kit Disulfide Bond Enhancer and SHuffle Competent E coli for heterologous in vitro and in vivo cellulase expression E6800
- Using the PURExpress In Vitro ProteinSynthesis Kit for Heterologous In Vitro Expression and Functional Screening of FMN dependent Oxidoreductase Variants E6800
Thoroughly mix solutions A and B before using. Do not vortex Solution B or ribosomes, mix gently.
Solution A may have a cloudy white appearance. Add to the reaction as a uniform suspension.
Assemble the reactions in the following order on ice: Solution A, Solution B, RNAse Inhibitor, Water, Template DNA or RNA
Once reaction is assembled take time to make sure everything is thoroughly mixed by gently pipetting up and down, pulse spin and place at 37C for 2 to 4 hours.