Standard Protein Synthesis Service

Bioneer’s Standard Protein Synthesis Service provides recombinant proteins within at least 1 week. This service uses an automatic protein synthesis system (ExiProgen™ Protein Synthesis System) to synthesize various proteins such as enzymes, growth factors, and hormones.

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Cat. No.

S-2500

Product Overview

Principle of Protein Synthesis Service using ExiProgen™ is shown below.

Add template DNA for target protein expression into protein synthesis kit and install in ExiProgen™ to get the purified protein.

The process of protein synthesis is in two steps.

Proteins are expressed by using E. coli cell-free protein expression and purified by Ni-affinity.

They are provided dissolved form in storage buffer.

Features and Benefits

Reduced experimental time

Delivery within 1 week through protein synthesis by E. coli cell-free protein expression and Ni-affinity purification
Advanced expression technologies
Synthesis of various proteins (enzymes, growth factors, hormones, antigens, antibodies, toxin proteins, etc.) difficult to be done in the existing in vivo technology

Workflow

▶ Schematics of service process

Type I) E. coli expression vectors (ex) T7-based expression vector)

Note) Available for gene of 0.3~4.0 kb in ORF (open reading frame)

Type II) Non-expression vectors (ex) Cloning vector/target gene)

Note) Available for gene of 0.3~1.5 kb in ORF (open reading frame).

▶ Example of protein synthesis

1 Normal proteins

M: AccuLadder ™ Protein Size Marker (Broad)
1: RNase H (Ribonuclease H),
2: DUSP3 (Dual specific phospatase 3),
3: CAT (Chloramphenicol acetyl transferase),
4: Poly A polymerase,
5: MMLV RTase (M-MLV reverse transcriptase)

M: AccuLadder ™ Protein Size Marker (Broad)
1: Calml3 (Calmoduline like protein),
2: hGH (Human Growth Hormone),
3: FGF2 (Fibroblast growth factor 2),
4: hLIF (Leukemia inhibitory factor)

2 Disulfide bonded proteins

M: AccuLadder ™ Protein Size Marker (Low)
1 : ScFv (Single-chain variable fragment, 2 disulfide bonds)
2 : rPA (recombinant plasminogen activator, 9 disulfide bonds)

* rPA synthesized through disulfide bond kit has enzymatic activity.
Activity assay using chromogenic substrate S-2288.

3 Tag-free proteins

M: AccuLadder ™ Protein Size Marker (Low),
1 : Expressed pBIVT-TEV-AcGFP
2 : Purified TEV-AcGFP (His-tagged)
3 : Final AcGFP (His tag-free sample)

4 Membrane proteins

M: AccuLadder ™ Protein Size Marker (Low)
1 : FLAP (35 kDa)
2 : bR (43 kDa)

* FLAP (5-lipoxygenase activating protein) has 4 transmembrane domain.
* bR (bacteriorhodopsin) has 7 transmembrane domains.

Literature/Support

◈ MSDS

• Standard Protein Synthesis Service MSDS

◈ Brochure

• All about Bioneer's Custom Services

◈ Quality Assurance

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

• ISO 9001 – certificate

Ordering Info

Amount of sample	E. coli expression vector¹⁾	> 30 µg (A_260/280 : 1.7 ~ 2.0, A_260/230 : >1.5)
	Non-expression vector²⁾	> 2 µg (A_260/280 : 1.7 ~ 2.0, A_260/230 : >1.5)
Cost of synthesis	$1,000
	Note) If protein synthesis fails, you will pay only set-up charge ($300).
Delivery form³⁾	1. Data report - E-mail 2. Protein - 100 µg (0.1~1.0 mg/ml of solution form)
Storage buffer	50 mM Tris-Cl (pH7.6), 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.05% NaN₃, 50% Glycerol
Period of synthesis⁴⁾	7 ~ 14 working days
Extra fee⁵⁾	$700/100 µg

¹⁾ If amount and purity of plasmid DNA are inconsistent with sample condition, an increase in quantity service of DNA will proceed and additional cost will be incurred.

²⁾ Non-expression vector means samples such as cloning vector of PCR Product. These samples require process of template DNA preparation for protein expression and
therefore additional cost ($200) will be charged.

³⁾ The 100 µg means amount of total protein in solution.

⁴⁾ The period may increase according to state of DNA sample or amount of protein and shipping period of sample is excluded from the period.

⁵⁾ The extra fee refers to the cost of synthesizing additional 100 µg of protein, and for those who do not provide 30ug of DNA sample, an addtional $50 will be charged.

▶ Quotation and order process

1 Download the order sheet from the link on web site.

2 Fill out the form and email the form to: export1@bioneer.com

* Please fill in the personal information columns (contact information). We need it to fine-tune the service and expedite the order.

3 We will review the order and send quote to you via email.

4 If you decide to go ahead with the service, please send the DNA sample to Bioneer global center.

Address: 71, Techno 2-ro, Yuseong-gu, Daejeon, Republic of Korea, 34013.

▶ Consultation

Contact
Tel. +82-42-939-6333 (Customer Support Center)
E-mail. sales@bioneer.co.kr or proteinorder@bioneer.com
Consultation Time
Weekdays from 9:00 am to 6:00 pm (closed on weekends and holidays)

FAQ

How can I order protein synthesis service?

After filling and sending the order form according [Quotation and order process] on the Ordering Information page, please send the template DNA samples according to the conditions below.

DNA type	*E. coli* expression vector(ex) pET etc T7-based vetor)	Non expression vector¹⁾ (ex) Cloning vector, PCR product)
DNA size²⁾	0.3 kb ~ 4.0 kb	0.3 kb ~ 1.5 kb
DNA amount	30 µg or more (A_260/280 : > 1.7~2.0, A_260/230 : >1.5)	2 µg or more (A_260/280 : > 1.7~2.0, A_260/230 : >1.5)

¹⁾ Additional cost of $100 will be charged if the sample is in the form of non-expression vector as these require process of template DNA preparation for protein expression

²⁾ DNA size represents the size of the ORF (Open Reading Frame) except the vector.

What should I do not have enough template DNA samples?

You can use DNA amplication service. but an additional cost of $50 per 30 µg DNA will be charged.

How long does it take for the protein synthesis?

Generally, the protein synthesis takes one day, but depending on the amount of requested protein, it may take a longer time.

The exact time can be checked through the quotation.

What is the available protein size for your service?

Our company has synthesis data for molecular weights of 10~150 kDa.

If your requested protein is out of range, a fee of $100 will be charged as additional experiments will be required.

If I do not have the template DNA, is this service unavailable for me?

If you don’t have the template DNA for your desired proteins, you may try using Gene to Protein service which not only synthesizes genes, but also provide protein synthesis service.

In this case, you must pay an additional cost for the gene expression.

Please refer to Gene to Protein Service order page for more details.

What is the composition of the buffer containing the protein sample? What if I want to change it?

Generally, the buffer composition for the protein delivery is as the following:

Normal Proteins: 50 mM Tris-Cl (pH7.6), 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.05% NaN₃, 50% Glycerol

Disulfide Proteins: 50 mM Tris-Cl (pH7.6), 100 mM NaCl, 1 mM DTT, 0.05% NaN₃, 50% Glycerol

If you want to recieve your proteins in a specific buffer, please state its name in the order form. However, please note that additional time and costs will be required.

What kinds of kits are used for protein synthesis?

The kits used in the protein synthesis services depends on the protein characteristics Refer to the following table:

Protein Type	Cat. No.	Product Description
Normal Protein	K-7300	ExiProgen EC Protein Synthesis Kit
Disulfide Protein	K-7330	ExiProgen EC-Disulfide Protein Synthesis Kit
His-tag free Protein	K-7320	ExiProgen EC-Tagfree Protein Synthesis Kit

Does the protein synthesis method differ depending on the template DNA?

In case of E. coli expression vector having T7 promoter, T7 terminator, RBS sequence and His-tag, protein synthesis can be done immediately without other steps.

On the other hand, in case of non-expression vector, there is no sequence that can undergo protein expression or purification. Thus, additional procedures must be done beforehand.

A two-step process is done after the primer design. First, a primer design for the gene amplification is performed and PCR is performed for procedure of making template DNA for protein expression. Then, a protein synthesis is done with the purified template DNA.