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ExiProgen™ EC-Bulk Protein Synthesis Kit

 Catalog NumberDescriptionPrice(USD) 
♦K-7340ExiProgen™ EC-Bulk Protein Synthesis Kit$2037.50add cart

Specfications

Shipping conditiondry ice and ice pack

Product Overview

ExiProgen™ EC-Bulk Protein Synthesis Kit allows to synthesize up to 10 mg of proteins using BIONEERs 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.

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

Components of kit

Box 1
(Protein Purification)		 Cartridge 96 well x 1 ea Store at
4 ~ 8 °C
Cartridge  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
Box 2
(Protein Expression)		 Cartridge 96 well x 1 ea Store at
-70 ~ -20 °C
Cartridge  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

 

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)

Price Competitiveness compared with other companies

Figures

1. Synthesis of fluorescent protein

Figure 1. Positive control protein synthesis data of ExiProgen™ EC-Bulk Protein 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

Literature/Support

Frequently asked questions (FAQs)

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.)

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

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: A260/280 : 1.7-2.0, A260/230 : >1.5 .

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.

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).