ExiProgen™ EC-Bulk Protein Synthesis Kit

ExiProgen™ EC-Bulk Protein Synthesis Kit allows to synthesize up to 10 mg of proteins using BIONEER’s patented SECF (Stepwise Exchange Cell-Free) technology.
This kit uses Expression protocol (No. 906) and Purification protocol(No.907). All materials required for protein expression & purification are included in the kit. The final protein products will be provided in an aqueous form dissolved in the storage buffer.

※ This product is shipped in dry ice and ice pack by cartridge

$1,630.00

Cat. No.

K-7340

Product Overview

Easy Bulk Protein Synthesis, ExiProgen™ EC Protein Synthesis Kit

Features and Benefits

Productive
Obtain up to 10 mg of purified protein per reaction.
Quick & Convenient
Synthesis your proteins without the need of cell culturing, which is a time-consuming process
Comprehensive
Synthesize proteins that are difficult to be done through in vivo experiments

Overview of experiment

Technical/Specs

▶ Components of kit

Box 1 (Protein Expression)	Cartridge ①	96 well x 1 ea	Store at -70 ~ -20 ℃
	Cartridge ②	96 well x 1 ea
	E. coli extract	6 mL x 1 bottle
	Master mix	10 mL x 1 bottle
	DEPC DW	6 mL x 1 bottle
Box 2 (Protein Purification)	Cartridge ①	96 well x 1 ea	Store at 4 ~ 8 ℃
	Cartridge ②	96 well x 1 ea
	Bulk SECF device	1 pack (2 ea/pack)
	Disposable filter tip	2 packs (12 ea/pack)
	Quick Manual	1 ea

Cartridges for protein expression
	*Feeding Buffer :Buffer for protein expression It contains NTPs, amino acids, and energy source.
Cartridges for protein purification
	Ni-NTA Magnetic bead :Beads for protein purification Binding/Washing buffer :Buffer for washing process to remove impurities Elution buffer :Buffer for purification or target proteins Storage buffer :Buffer for storage of final target proteins (50mM Tris-Cl (pH7.6), 100mM NaCl, 1mM DTT, 0.05%(v/v) NaN3, 50%(v/v) Glycerol)

▶ Experimental Data

1. Synthesis of fluorescent protein

Figure 1. Positive control protein synthesis data of ExiProgen™EC-BulkProtein Synthesis Kit
Plasmid DNA; pBIVT-AcGFP used, PCR Product; AcGFP PCR (2F/2R primer of K-7400 kit) Product used.
NC; No-DNA sample, M; AccuLadder™ Protein size marker (Low) (Bioneer Cat.No. D-2020),
E; Expression sample, U; Unbound sample, W; Washing sample, S; Storage sample
B; Bead sample, B(S); Supernatant of bead sample

2. Synthesis of various proteins

Figure 2. High-value protein synthesis data of ExiProgen™ EC-Bulk Protein Synthesis Kit
M; AccuLadder™ Protein size marker (Low) (Cat. No. D-2020, Bioneer),
E; Expression sample, U; Unbound sample, W; Washing sample, S; Storage sample

◈ Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001 - certificate

References

▶ List of theses using Bioneer ExiProgen™

Issue year	Title of Thesis	Author	Thesis information
2018	Highly efficient genome editing by CRISPR-Cpf1 using CRISPR RNA with a uridinylate-rich 3′-overhang	Bin Moon S, Lee JM, Kang JG, Lee NE, Ha DI, Kim DY, Kim SH, Yoo K, Kim D, Ko JH, Kim YS	Nat Commun. 2018 Sep 7;9(1):3651.
2018	Receptor-mediated dimerization of JAK2 FERM domains is required for JAK2 activation.	Ryan D Ferrao, Heidi JA Wallweber, Patrick J Lupardus	*Elife.* 2018 Jul 25;7.
2018	Selectively Modulating Conformational States of USP7 Catalytic Domain for Activation.	Özen A, Rougé L, Bashore C, Hearn BR, Skelton NJ, Dueber EC	*Structure.* 2018 Jan 2;26(1):72-84.
2016	The OsCYP19-4 Gene Is Expressed as Multiple Alternatively Spliced Transcripts Encoding Isoforms with Distinct Cellular Localizations and PPIase Activities under Cold Stress.	Lee A, Lee SS, Jung WY, Park HJ, Lim BR6 Kim HS, Ahn JC, Cho HS	*Int J Mol Sci.* 2016 Jul 19;17(7).
2016	The role of MscL amphipathic N terminus indicates a blueprint for bilayer-mediated gating of mechanosensitive channels.	Bavi N, Cortes DM, Cox CD, Rohde PR, Liu W, Deitmer JW, Bavi O, Strop P, Hill AP, Rees D, Corry B, Perozo E, Martinac B	*Nat Commun.* 2016 Jun 22;7:11984.
2016	Removal of the mechanoprotective influence of the cytoskeleton reveals PIEZO1 is gated by bilayer tension.	Cox CD, Bae C, Ziegler L, Hartley S, Nikolova-Krstevski V, Rohde PR, Ng CA, Sachs F, Gottlieb PA, Martinac B	*Nat Commun.* 2016 Jan 20;7:10366.
2016	Overexpression of OsCYP19-4 increases tolerance to cold stress and enhances grain yield in rice (Oryza sativa L.)	Yoon DH, Lee SS, Park HJ, Lyu JI, Chong WS, Liu JR, Kim BG, Ahn JC, Cho HS	*J Exp Bot.* 2016 Jan;67(1):69-82.
2015	Development of a Recombinant Protein Vaccine Based on Cell-Free Protein Synthesis for Sevenband Grouper Epinephelus septemfasciatus Against Viral Nervous Necrosis.	Kim JO, Kim JO, Kim WS, Oh MJ	*J Microbiol Biotechnol.* 2015 Oct;25(10):1761-7.

Ordering Info

▶ Ordering Information

Cat. No.	Product Description
K-7340	ExiProgen™ EC-Bulk Protein Synthesis Kit

▶ Related Products

Cat. No.	Product Description
K-7300~02	ExiProgen™ EC Protein Synthesis Kit, 16, 32, 96 reactions
K-7350	pBVIT Vector Set-1
K-7400	ExiProgen™ ProXpress PCR Template Kit, 16 reactions
K-2633	AccuPower^® ProFi Taq PCR PreMix (96 T, 50 μl)
K-3037	AccuPrep^® PCR/Gel DNA Purification Kit (200 reactions)
D-2010	AccuLadder™ Protein size marker (Broad), 500 μl
A-5041	ExiProgen™

FAQ

What is the range of protein sizes that can be synthesized?

The recommended protein size is from 10 kDa to 150 kDa.
Synthesis of proteins that does not fall in the size range may be possible, but is not guaranteed.
(Please note that the cut-off size of the Bulk SECF device included in the kit is 6 to 8 kDa. Thus, proteins smaller than 10kDa may be lost during the expression and purification process of the instrument.)

How long does the protein synthesis take?

There are separate protocols for protein expression and purification. While the former, No. 906 [Bulk_Expression], takes about 25 hours, the latter, No. 907 [Bulk_Purification], takes about 15 hours.
We recommend to set the protocols efficiently like below.
(Mon) 3:00 ~ 4:00 P.M.: Start the protein expression step → (Tue) 4~5:00 P.M.: Collect expressed protein sample and start the protein purification step → (Wed) 9:00 A.M : Collect the purified protein sample

How do I prepare the template DNA?

Template DNA must be constructed as the following:
[T7 promoter-RBS-Target gene(with His-tag)-T7 terminator]
Both plasmid DNA and PCR products can be used.

1) Plasmid DNA
pBIVT vector set-1 (Cat. No. K-7350) is optimized to in vitro transcription and translation.
Other expression vectors having the above structures can be also used.

2) PCR product type
A. Non-expression Vector / Genomic DNA
e.g.) T-vector
you may use ExiProgen™ ProXpress PCR Template Kit (Cat. No. K-7400, Bioneer), which allows to synthesize templates without undergoing the cloning process.
B. Expression Vector that require amplification
: You may use our 2F/2R Primer set (Cat. No. K-7400, Bioneer)and AccuPower^® ProFi Taq PCR PreMix (Cat. No. K-2633, Bioneer).

We recommend to use plasmid DNA rather than PCR products when the target protein size is more than 50 kDa.
Also, please prepare DNA with the following purity: A_260/280 : 1.7-2.0, A_260/230 : >1.5 .

Can the protein expression be affected by template DNA sequences?

Yes. As our protein synthesis kits are based on E. coli expression systems, we recommend to undergo codon optimization process based on E. coli.
The expression level of proteins will be better for the samples with E. coli-based codon optimized sequences.

What is the best possible way to get the maximum amount of protein?

Finding the optimal amount of template DNA is the most important step. Therefore, before using this kit, we recommend trying to undergo screening protocol using ExiProgen™ EC Protein Synthesis Kit (K-7300~02).

If you have any other questions, please contact sales@bioneer.com

ExiProgen™ EC-Bulk Protein Synthesis Kit

Features and Benefits

Overview of experiment

▶ Components of kit

▶ Experimental Data

▶ Price Competitiveness compared with other companies

◈ Manual

◈ MSDS

◈ Flyer

◈ Quality Assurance

▶ List of theses using Bioneer ExiProgen™

▶ Ordering Information

▶ Related Products