PUREfrex Reaction Supplements - supplements expand the potential of base kits -

Chaperones / Forming disulfide bonds / Other proteins

Overview

Depend on your protein’s property, we have supplements to add to PUREfrex® kit. Molecular chaperones, DnaK Mix or GroE Mix, are for improving folding and solubility. DsbC Set or PDI Set are for making disulfide bonds. Depending on each protein, adding those supplements to PUREfrex® kit helps effective translation and folding simultaneously.

Chaperones　 -DnaK Mix / GroE Mix-

Molecular chaperones such as hsp70 (e.g. E. coli DnaK) assist the correct folding of the newly synthesized proteins. Because PUREfrex® does not contain any molecular chaperones, some proteins may have difficulty in forming the correct conformation and be insoluble after synthesis. In such cases, molecular chaperones could contribute to make them soluble and active.

　

PUREfrex Reaction Supplements - Chaperones - Ordering Information

Product Name	Catalog Number	Total Reaction Volume
DnaK Mix	GFK-PF003-0.5-EX	500 ul
	GFK-PF003-10ML-EX	10 ml（5 x 2 ml）
	GFK-PF003-50ML-EX	50 ml（25 x 2 ml）
GroE Mix	GFK-PF004-0.5-EX	500 ul
	GFK-PF004-10ML-EX	10 ml（5 x 2 ml）
	GFK-PF004-50ML-EX	50 ml（25 x 2 ml）

Experiment Workflow

read more

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 DnaK Mix／GroE Mix as supplement to PUREfrex® reaction mixture if the solubility or activity of the synthesized protein is low.

DnaK Mix

DnaK Mix is newly developed supplements for PUREfrex® series (Cell-free protein Synthesis Kits) to assist proper folding and improving solubility of the protein that needs molecular chaperones for proper protein folding, since PUREfrex® series (Cell-free protein Synthesis Kits) do not contain molecular chaperones.

DnaK Mix is consisted of highly purified DnaK, DnaJ and GrpE from E.coli with optimized ratio. DnaK known as Hsp70 has ATPase activity and is stimulated by co-chaperones, DnaJ and GrpE. DnaJ facilitates the ATPase activity of DnaK and could bind to a hydrophobic region of synthesized protein. GrpE stimulates ADP/ATP exchange rate of DnaK.

DnaK Mix works very well with PUREfrex® series (Cell-free protein Synthesis Kits) and DsbC Set (Supplement for forming disulfide bonds) in a same tube for protein synthesis reaction, which could lead to the preparation of your protein in proper folding with good solubility according to the character of your protein.

Figure. Effect of DnaK Mix on the activity of synthesized luciferase.

DnaK Mix Kit Contents and Storage

Reagent	Quantity　for 500 μL rxn	Description	Storage ^*1
DnaK Mix	25 µL	100µM DnaK, 20µM DnaJ and 20µM GrpE from E.coli in 30% glycerol buffer (No tags)	-80 °C ^*2
Dilution buffer	500 µL	30% glycerol buffer	-20 °C

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

Protocol

read more

The amounts given are for 20 µL reaction with PUREfrex®2.0. For scaling up the reaction, adjust the volume of reagents accordingly.

• 1)Thaw Solution I by incubation at room temperature or 37 °C for 1 minute for completely dissolving, and then cool on ice.
• 2) Thaw Solution II, III and DnaK Mix on ice.
• 3) Mix Solution I, II, III and DnaK Mix 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.
• 5) Incubate the tube at 37 °C for 2 - 6 hours on a heat block or a water bath.
• 6) Analyze the synthesized products. Please add the same amount of H2O to the reaction for the sample of SDS-PAGE.

Contents of a standard PUREfrex reaction	volume
Water	6-X μL
Solution I	10 μL
Solution II	1 μL
Solution III	2 μL
DnaK Mix^*	1 μL
Template DNA	X μL
Total	20 μL
^* The optimum concentration of DnaK Mix depends on a protein. Please use "Dilution buffer" (in the kit) to dilute the DnaK mix.

GroE Mix

GroE Mix is a supplements for PUREfrex® series (Cell-free protein Synthesis Kits) to assist proper folding and improving solubility of the protein that needs molecular chaperones for proper protein folding, since PUREfrex® series (Cell-free protein Synthesis Kits) do not contain molecular chaperones.

GroE Mix is constituted of highly purified GroEL (known as Hsp60) and GroES (working in conjunction with GroEL) from E.coli with the optimized ratio.

GroE Mix works very well with PUREfrex® series (Cell-free protein Synthesis Kits) in a same tube for protein synthesis reaction, which could lead to the preparation of your protein in proper folding with good solubility according to the character of your protein.

Figure. Effect of GroE Mix on the solubility of synthesized proteins.
(T: total reaction mixture, S: supernatant, P: precopitate fractions)

GroE Mix Kit Contents and Storage

Reagent	Quantity　for 500 μL rxn	Description	Storage ^*1
Water	6-X μL
GroE Mix	12.5 µL	20µM GroEL, 14-mer form, and 40µM GroES, 7-mer form, from E.coli in 30% glycerol buffer (No tags)	-80 °C ^*2
Dilution buffer	500 µL	30% glycerol buffer	-20 °C

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

Protocol

read more

The amounts given are for 20 µL reaction with PUREfrex®2.0. For scaling up the reaction, adjust the volume of reagents accordingly.

• 1) Thaw Solution I by incubation at room temperature or 37 °C for 1 minute for completely dissolving, and then cool on ice.
• 2) Thaw Solution II, III and GroE Mix on ice.
• 3) Mix Solution I, II, III and GroE Mix by vortex and centrifuge briefly to collect each solution at the bottom.
• 4) Assemble the reaction mixture in a tube as follows. DO NOT add GroE Mix at this step. Add the template DNA to
• 5) Incubate the tube at 37 °C for 15 minutes.
  (To prevent an inhibition of transcription, we recommend a pre-incubation before addition of GroE Mix.)
• 6) Make a Two-fold dilution of GroE*1 Mix with Dilution buffer and add 1 µL to incubated tube in Step 5.
  (The optimum concentration of GroE Mix depends on a protein. Please use "Dilution buffer" (in the kit) to dilute the GroE mix.)
• 7) Incubate the tube at 37 °C for 2 - 6 hours on a heat block or a water bath.
• 8) Analyze the synthesized products. Please add the same amount of H2O to the reaction for the sample of SDS-PAGE.

GroE Mix Kit Contents and Storage

Contents of a standard PUREfrex reaction	volume
Water	6-X μL
Solution I	10 μL
Solution II	1 μL
Solution III	2 μL
Template DNA	X μL
Total	19 μL

Forming disulfide bonds 　 -DsbC Set / PDI Set-

Formation of disulfide bonds is important for folding and stability of secretory proteins such as enzymes or antibodies. Disulfide bonds are usually formed by oxidation of sulfhydryl groups (SH-) of adjacent cysteine residues. Therefore, disulfide bond formation efficiency depends on redox state. Additionally, disulfide bond isomerase which can catalyze the exchange of disulfide bridges may be required for correct cysteine pairing.

　

PUREfrex Reaction Supplements - Forming disulfide bonds - Ordering Information

Product Name	Catalog Number	Total Reaction Volume
DsbC Set	GFK-PF005-0.5-EX	500 ul
	GFK-PF005-10ML-EX	10 ml（5 x 2 ml）
	GFK-PF005-50ML-EX	50 ml（25 x 2 ml）
PDI Set	GFK-PF006-0.5-EX	500 ul
	GFK-PF006-10ML-EX	10 ml（5 x 2 ml）
	GFK-PF006-50ML-EX	50 ml（25 x 2 ml）

Experiment Workflow

read more

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 DsbC Set／PDI Set as supplement to PUREfrex® reaction mixture if the solubility or activity of the synthesized protein is low.

DsbC Set

DsbC Set is the supplement by adding to PUREfrex® series (Cell-free protein Synthesis Kits) for synthesizing proteins containing a disulfide bond in an active form.

Formation of a disulfide bond is an important process for folding and stability of secretory proteins such as enzymes or antibodies. A disulfide bond is usually formed by the oxidation of sulfhydryl group (SH-) of adjacent cysteine residues. Therefore, the formation efficiency of a disulfide bond depends on redox state. Additionally, disulfide bond isomerase which can catalyze the exchange of disulfide bridges may be also required for a correct pairing of cysteines.

DsbC Set is constituted of highly purified DsbC from E.coli known as disulfide bond isomerase, which can catalyze the disulfide bridge exchange, and GSSG (oxidized Glutathione) to make an oxidized environment.

　

DsbC Set Kit Contents and Storage

Reagent	Quantity　for 500 μL rxn	Description	Storage ^*1
GSSG	25 µL	Oxidized glutathione (60 mM)	-20 °C
DsbC	25 µL	E.coli DsbC (320 µM *2) (No tags)	-80 °C ^*3
Dilution buffer	500 µL	30% glycerol buffer	-20 °C

• ※1) Indicated temperature shows storage temperature after opening the kit. Please store the kit at -80 °C before opening.
• ※2) It's same concentration as 7.5 mg/mL described in former instruction.
• ※3) The rest of solution should be frozen rapidly in liquid nitrogen or dry ice/ethanol, and be stored at -80 °C. Divide into aliquots, if necessary, and avoid refreeze and thaw as much as possible.

Protocol

read more

This is a standard protocol for synthesizing proteins containing disulfide bonds.
Each solution of DsbC Set and PUREfrex®2.1 are mixed together in a same tube.

• 1) Thaw Solution I, Cysteine, GSH and GSSG by incubation at room temperature or 37 °C for 1 minute completely, and then cool on ice.
• 2) Thaw Solution II, III and DsbC 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.
• 5) Incubate the tube at 37 °C for 2 - 6 hours with heat block or water bath.
  Protein synthesis reaction is almost done until 6 hours, but some proteins require longer incubation (e.g. 24 hours) to form disulfide bonds between the correct pair of cysteine residues.
• 6) Analyze the synthesized products. Please add the same amount of H2O to the reaction for the sample of SDS-PAGE.

Contents of a standard PUREfrex reaction	volume
Water	5-X μL
Solution I^*1	8 μL
10 mM Cysteine	1 μL
80 mM GSH	1 μL
60 mM GSSG	1 μL
Solution II	1 μL
Solution III	2 μL
DsbC (80 μM) ^*2	1 μL
Template DNA^*3	X μL
Total	20 μL

• ※1) This volume is different from Solution I of PUREfrex®2.0.
• ※2) DsbC is diluted with Dilution buffer included in DsbC Set.
• ※3) Please visit the template DNA preparation site.

PDI Set

PDI Set is the supplement by adding to PUREfrex® series (Cell-free protein Synthesis Kits) for synthesizing proteins containing a disulfide bond in an active form.

Formation of a disulfide bond is an important process for folding and stability of secretory proteins such as enzymes or antibodies. A disulfide bond is usually formed by the oxidation of sulfhydryl group (SH-) of adjacent cysteine residues. Therefore, the formation efficiency of a disulfide bond depends on redox state. Additionally, disulfide bond isomerase which can catalyze the exchange of disulfide bridges may be also required for a correct pairing of cysteines.

PDI Set includes oxidized glutathione (GSSG), human PDI (protein disulfide isomerase) and human Ero1α (ER oxidoreductin-1 to reoxidize PDI).

PDI Set Kit Contents and Storage

Reagent	Quantity　for 500 μL rxn	Description	Storage ^*1
GSSG	25 µL	Oxidized glutathione (60 mM)	-20 °C
PDI	25 µL	Human protein disulfide isomerase with no tags (200 µM)	-80 °C ^*2
Ero1α	25 µL	Human ER oxidoreductin-1 to reoxidize PDI with no tags (5 µM)	-80 °C ^*2
Dilution buffer	500 µL	30% glycerol buffer	-20 °C

• ※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.

Protocol

read more

＜CASE 1＞ PDI and GSSG

This is a standard protocol for synthesizing proteins containing disulfide bonds using PDI Set 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, 1 mM GSSG and 10 µM PDI.

• 1) Thaw Solution I, Cysteine, GSH and GSSG by incubation at room temperature or 37 °C for 1 minute completely, and then cool on ice.
• 2) Thaw Solution II, III and PDI on ice.
• 3) Mix each solution by vortex and centrifuge briefly to collect each solution at the bottom.
• 4) Dilute GSSG 3-fold with water.
• 5) Assemble the reaction mixture in a tube as follows. Add the template DNA to 1 - 3 ng/µL per 1 kb.
  "PDI Set" is the supplement by adding to PUREfrex® for synthesizing proteins containing a disulfide bond in an active form.
• 6) Incubate the tube at 37 °C for 2 - 6 hours with heat block or water bath.
  Protein synthesis reaction is almost done until 6 hours, but some proteins require longer incubation (e.g. 24 hours) to form disulfide bonds between the correct pair of cysteine residues.
• 7) Analyze the synthesized products. Please add the same amount of H2O to the reaction for the sample of SDS-PAGE.

PDI Set Kit Contents and Storage

Contents of a standard PUREfrex reaction	volume
Water	5-X μL
Solution I^*1	8 μL
10 mM Cysteine	1 μL
80 mM GSH	1 μL
60 mM GSSG^*2	1 μL
Solution II	1 μL
Solution III	2 μL
PDI	1 μL
Template DNA^*3	X μL
Total	20 μL

• ※1) This volume is different from Solution I of PUREfrex®2.0.
• ※2) Please use 3-fold diluted GSSG. Standard final concentration of GSSG is 1 - 3 mM with 4 mM GSH (reduced glutathione) in PUREfrex®2.1. We recommend to check the optimal concentration of GSSG because it depends on the target protein and the kind and concentration of reducing agent.
• ※3) Please visit the template DNA preparation site.

＜CASE 2＞ PDI and Ero1α

This is a standard protocol for synthesizing proteins containing disulfide bonds using PDI Set 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, 10 µM PDI and 0.25 µM Ero1α.

• 1) Thaw Solution I, Cysteine and GSH by incubation at room temperature or 37 °C for 1 minute for completely dissolving, and then cool on ice.
• 2) Thaw Solution II, III, PDI and Ero1α 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.
• 5) Incubate the tube at 37 °C for 2 – 6 hours with heat block or water bath.
  Protein synthesis reaction is almost done until 6 hours, but some proteins require longer incubation (e.g. 24 hours) to form disulfide bonds between the correct pair of cysteine residues.
• 6) Analyze the synthesized products. Please add the same amount of H2O to the reaction for the sample of SDS-PAGE.

PDI Set Kit Contents and Storage

Contents of a standard PUREfrex reaction	volume
Water	5-X μL
Solution I^*1	8 μL
10 mM Cysteine	1 μL
80 mM GSH	1 μL
Solution II	1 μL
Solution III	2 μL
200 μM PDI	1 μL
5 µM Ero1α^*2	1 μL
Template DNA^*3	X μL
Total	20 μL

• ※1) This volume is different from Solution I of PUREfrex®2.0.
• ※2) Standard final concentration of PDI/Ero1α is 1/0.025 - 10/0.25 µM. We recommend to check the optimal concentration of PDI and Ero1α because it depends on the target protein. Please use attached Dilution Buffer for diluting PDI and Ero1α.
• ※3) Please visit the template DNA preparation site.

Other proteins 　-EF-P-

The E. coli translation factor EF-P (elongation factor P), a bacterial homolog of eukaryotic initiation factor 5A (eIF5A), functions to promote peptide bond formation between the first methionine and the next amino acid as well as the elongation of consecutive proline residues (3 or more residues). In bacteria, LYS34 of EF-P is modified post translationally to ß-lysllysine by EpmA (YjeA/GenX) and EpmB (YjeK). This modification is important to EF-P activity. PUREfrex EF-P Supplement contains this modification at LYS34.

PUREfrex Reaction Supplements - EF-P- Ordering Information

Product Name	Catalog Number	Total Reaction Volume
EF-P	GFK-PF052-0.5-EX	500 ul
	GFK-PF052-10ML-EX	10 ml（5 x 2 ml）
	GFK-PF052-50ML-EX	50 ml（25 x 2 ml）
This is supplement for synthesizing proteins containing disulfide bonds in an active form.

Experiment Workflow

read more

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.

EF-P

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.

Lysine at 34th of EF-P is post-translationally modified to β-lysllysine by EpmA (YjeA/GenX) and EpmB (YjeK) and the modification is important for the activity.

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

EF-P Set Kit Contents and Storage

Reagent	Quantity　for 500 μL rxn	Description	Storage ^*1
EF-P	12.5 µL	EF-P with no tags (40 µM)	-80 °C ^*2
Dilution buffer	500 µL	30% glycerol buffer	-20 °C

• ※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.

Protocol

read more

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.
• 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.

EF-P Set Kit Contents and Storage

Contents of a standard PUREfrex reaction	volume
Water	6.5-X μL
10 mM Cysteine	1 μL
80 mM GSH	1 μL
Solution II	1 μL
Solution III	2 μL
40 μM EF-P	0.5 μL
Template DNA^*2	X μL
Total	20 μL

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

PUREfrex Ordering Information

PUREfrex Ordering Information

Product Name (click for price and order info)	Catalog Number	Total Reaction Volume	Notes
Entry Model Kits
PUREfrex® 2.0 mini	GFK-PF201-0.1-EX	100 μL	New item !
PUREfrex® 2.0 mini is an entry model designed for those who are trying protein synthesis for the first time with a cell-free system.Product Information Page：PUREfrex® 2.0 mini
Kits
PUREfrex® 2.0	GFK-PF201-0.25-EX	250 ul
	GFK-PF201-0.25-5-EX	5 x 250 ul (packaged as 5 x 250 ul)	Sales to be discontinued
	GFK-PF201-2ML-EX	1 x 2 ml
	GFK-PF201-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF201-10-EX
	GFK-PF201-50ML-EX	25 x 2 ml	New size !
Current generation kit for protein synthesis. Contains DTT as a reducing agent. Product Information Page：PUREfrex® 2.0
PUREfrex® 2.1	GFK-PF213-0.25-EX	250 ul
	GFK-PF213-0.25-5-EX	5 x 250 ul (packaged as 5 x 250 ul)	Sales to be discontinued
	GFK-PF213-2ML-EX	1 x 2 ml	New size!
	GFK-PF213-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF213-10-EX
	GFK-PF213-50ML-EX	25 x 2 ml	New size !
Current generation kit for proteins with disulfide bonds. Formulated for user control of redox environment. Product Information Page：PUREfrex® 2.1
PUREfrex® 1.0	GFK-PF001-0.25-EX	250 ul
	GFK-PF001-0.25-5-EX	1250 ul (packaged as 5 x 250 ul)	Sales to be discontinued
	GFK-PF001-2ML-EX	1 x 2 ml	New size!
	GFK-PF001-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF001-10-EX
	GFK-PF001-50ML-EX	25 x 2 ml	New size !
Prior generation kit for protein science research and synthetic biology. Components are fully disclosed. Product Information Page：PUREfrex® 1.0
Custom formulated kits are based on 1.0 components. Please inquire about PUREfrex® custom.
PUREfrex Reaction Supplements
DnaK Mix	GFK-PF003-0.5-EX	for 500 ul of reaction mix
	GFK-PF003-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF003-10-EX
	GFK-PF003-50ML-EX	25 x 2 ml	New size !
GroE Mix	GFK-PF004-0.5-EX	for 500 ul of reaction mix
	PF004-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF004-10-EX
	PF004-50ML-EX	25 x 2 ml	New size !
Supplements for synthesis of aggregate-prone proteins. Product Information Page：PUREFrex Reaction Supplements
DsbC Set	GFK-PF005-0.5-EX	for 500 ul of reaction mix
	PF005-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF005-10-EX
	PF005-50ML-EX	25 x 2 ml	New size !
PDI Set	GFK-PF006-0.5-EX	for 500 ul of reaction mix
	PF006-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF006-10-EX
	PF006-50ML-EX	25 x 2 ml	New size !
Supplements for synthesis of proteins containing disufide bonds.. Product Information Page：PUREFrex Reaction Supplements
EF-P	GFK-PFS052-0.5-EX	for 500 ul of reaction mix
	PFS052-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PFS052-10-EX
	PFS052-50ML-EX	25 x 2 ml	New size !
Supplement for use when consecutive prolines are in target protein.  Product Information Page：PUREFrex Reaction Supplements

Notes

PUREfrex® is developed for in vitro research only. PUREfrex® 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”