#PF004-0.5-EX
For 500 µL Reaction
PUREfrex® is NOT included.

Store at -80ºC before opening

1. Overview

2. About PUREfrex®

GroE Mix is a newly developed supplement of PUREfrex®
to assist proper folding and solubility of your protein.
PUREfrex® is a cell-free protein synthesis reagent but has
no molecular chaperones (See next page).
When your protein of interest needs molecularchaperones for proper protein folding, GroE Mix could bea solution for that.
GroE Mix is constituted of highly purified GroEL (knownas Hsp60) and GroES (working in conjunction with GroEL)from E.coli with the optimized ratio.
GroE Mix works very well with PUREfrex®
(#PF001-0.25-EX or #PF201-0.25-EX) in a single tube forprotein synthesis reaction, which could lead to thepreparation of your protein in proper folding with goodsolubility.

PUREfrex® is a reconstituted cell-free protein synthesis
kit which GeneFrontier has developed based on the PURE
system technology. The PURE system is a cell-free protein
synthesis system, which has originally developed byProfessor Takuya Ueda at the University of Tokyo, and isc o n s i s t e d o f o n l y p u r i fi e d f a c t o r s n e c e s s a r y f o rtranscription, translation and energy regeneration (Ref. 1).
The target protein is synthesized by adding the template
DNA (or mRNA) to the reaction mixture. PUREfrex® is
consisted of only purified factors, therefore it enables toadjust the composition of the reaction mixture.
PUREfrex® is raised in the purity by improving thepreparation methods of ribosomes, tRNAs and all proteinsin the reaction mixture compared with the original PURE
system (Ref. 2). As the result, the contaminating
lipopolysaccharide from E. coli is reduced below 0.1 EU per
1 µL of reaction and other contaminants, such as RNase
and β-galactosidase, are also reduced.
In the PUREfrex®, all proteins have no tags for
purification or detection, therefore the target protein
would be synthesized and purified by any tag.

May 2023References) 1. Shimizu et al. (2001) Nat. Biotecnol., vol. 19, p. 751

SHARP Kashiwa Building, 4F
273-1 Kashiwa, Kashiwa-shi,
Chiba 277-0882 Japan

GeneFrontier Corporation
www.genefrontier.com

2. Shimizu et al. (2005) Methods, vol. 36, p. 299

GroE Mix is worked with PUREfrex® (#PF001-0.25-EX,#PF201-0.25-EX) in one tube. For example, 20 µL ofreaction is assembled as below.

12.5 µL

20µM GroEL and 40µM GroES in 30% glycerol buffer*1
Store at -80ºC

5. Incubate the tube at 37ºC for 15 minutes.*3
6. Make a Two-fold dilution of GroE Mix with Dilution*4

Buffer and add 1 µL to incubated tube in Step5.

*2

7. Incubate the tube at 37ºC for 2-4 hours.

Dilution Buffer (Clear)or administration to animals including human and shouldnot be used as food or cosmetics etc.
To avoid the contamination of nuclease, nuclease-freetreated water, reagents and materials should be used. We
also recommend wearing gloves and mask.
For information concerning commercial use of GroE Mix,
please contact GeneFrontier.
e-mail : purefrex@genefrontier.com【Outside Japan】【JAPAN】

Carlsbad, CA 92010, USA

KOTO-KU. TOKYO 135-0016, JAPAN2792 Loker Ave West, Suite 101
email: info@cosmobiousa.com

Phone/FAX: (+1) 760-431-4600
URL: www.cosmobiousa.com

Store at -80ºC before opening

GroE Mix (Purple)

GroE Mix is developed for in vitro research use only.
GroE Mix should not be used for the therapy, diagnostic

500 µL

1. Thaw Solution I by incubation at 30ºC for 1 minute,and then cool on ice.

8. Analyze the synthesized product.

2. Thaw Solution II, III and GroE Mix on ice.

30% glycerol bufferStore at -20ºC*1)
GroEL 14-mer and GroES 7-mer forms*2)
For storage at -80ºC, the rest of solution should be frozen rapidly inliquid nitrogen or dry ice/ethanol. Divide into aliquots, if necessary,and avoid refreeze and thaw as much as possible.

3. Mix Solution I, II, III and GroE Mix by vortex andcentrifuge 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 0.5-3 ng/µL per 1 kbp)

Water
Solution I
Solution II
Solution III
Template DNATotal#PF0018-X µL
10 µL
1 µL
1 µL
X µL
19 µL

#PF201
7-X µL
10 µL
1 µL
2 µL
X µL
19 µL*3)
To prevent an inhibition of transcription, we recommend a
pre-incubation before addition of GroE Mix.
*4)
The optimum concentration of GroE Mix depends on protein ofinterest. Please use Dilution buffer for dilution of GroE Mix.

TOYO EKIMAE BLDG. 2-20, TOYO 2-CHOME,
Phone: +81-3-5632-9610
FAX: +81-3-5632-9619

URL: https://www.cosmobio.co.jp/