EF-P

Ordering through the online e-commerce system is available only to customers in the USA and Canada.

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International sales of GeneFrontier's products are carried out by Cosmo Bio USA.
To request a quote or place an order, please contact Cosmo Bio USA.

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[Information]

• Exciting New Release! 50 mL Scale PUREfrex^® Kits

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Overview

“EF-P” is a supplement by adding to PUREfrex®series, cell-free protein synthesis kits.

EF-P (elongation factor P) is one of translation factors in E. coli and a homolog of eukaryotic initiation factor 5A (eIF5A). EF-P has a promoting activity for the peptide bond formation between the first methionine and the second amino acid and the elongation of consecutive proline residues (3 or more prolines).

Lysine at 34th of EF-P is post-translationally modified and the modification is important for the activity. “EF-P” kit includes post-translationally modified EF-P from E.coli.

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MS1 Extracted Ion Chromatogram of the isolated recombinant EF-P from E.coli.

Experiment Workflow

General procedures for using PUREfrex® to synthesize functional proteins are shown in Figure.

Start by preparing the template DNA suitable to the protein synthesis with PUREfrex® and examine whether the protein of interest is synthesized. Redesign the nucleotide sequence of the template DNA if too little protein is synthesized.

Add EF-P as supplement to PUREfrex® reaction mixture if the amout of synthesized protein is low and it contains consecutive proline residues.

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Protocols

This is a standard protocol for synthesizing proteins using EF-P and PUREfrex®2.1. For example, please assemble 20 µL of reaction mixture as below, in which the final concentration of each reagent is 0.5 mM Cysteine, 4 mM GSH and 1 µM EF-P.

1. Thaw Solution I, Cysteine and GSH by incubation at room temperature or 37 °C for 1 minute completely dissolving, and then cool on ice.


2. Thaw Solution II, III and EF-P on ice.


3. Mix each solution by vortex and centrifuge briefly to collect each solution at the bottom.


4. Assemble the reaction mixture in a tube as follows. Add the template DNA to 1 - 3 ng/µL per 1 kb.

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5. Incubate the tube at 37 °C for 2 - 6 hours with heat block or water bath.
   


6. Analyze the synthesized products. Please add the same amount of H₂O to the reaction for the sample of SDS-PAGE.

1. This volume is different from Solution I of PUREfrex®2.0.
2. Please visit the template DNA preparation site.

Specifications

Kit components   <EF-P>

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1. Indicated temperature shows storage temperature after opening the kit. Please store the kit at -80 °C before opening.
2. For storage at -80 °C, the rest of solution should be frozen rapidly in liquid nitrogen or dry ice/ethanol. Please divide into aliquots, if necessary, and avoid refreeze and thaw as much as possible.

Datasheet

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SDS

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Note

EF-P is developed for in vitro research only. EF-P should not be used for the therapy, diagnostic or administration to animals including human and should not be used as food or cosmetics etc. To avoid the contamination of nuclease, nuclease-free-treated water, reagents and materials should be used. We also recommend wearing gloves and mask.

"PUREfrex® is Registered in U.S. Patent and Trademark Office"

Applications

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On the Resources page, you can find posters and technical notes about PUREfrex®.
Additionally, there are papers that use PUREfrex® or PURE system technology.

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Related Products

When your protein needs an assistance of molecular chaperone or a formation of a disulfide bond to form a correct conformation, we have supplemental reagents by simply adding to PUREfrex® series.

Cell-free Protein Synthesis Kits

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Supplements – Chaperones –

These are supplement for PUREfrex® which is molecular chaperone to be aggregate-prone protein in a soluble form.

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Supplements – Forming disulfide bonds –

This is supplement for synthesizing proteins containing disulfide bonds in an active form.

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