ExiProgen™ EC-Disulfide Protein Synthesis Kit


ExiProgen™ EC-Disulfidee Protein Synthesis Kit enables it to automatically synthesize target protein with disulfide bonds. It can be much more effectively used than cell based protein expression system because it can synthesizes protein with up to nine disulfide bonds.


This kit is used for synthesizing a protein that contains disulfide bonds using ExiProgen™. Used in conjunction with ExiProgen™, it can synthesize a protein that contains up to nine disulfide bonds in a fully automated manner and more efficiently than using cell-based synthesis. After adding template DNA and associated reagents into ExiProgen™ and using protocol No. 905, the target protein can be synthesized within a day. All processes, including protein synthesis and dialysis, are performed in an automated manner and the final protein is harvested in any storage buffer of choice.

Features and Benefits

Protein synthesis with disulfide bond
This kit is used to synthesize a protein that has two or more disulfide bonds, providing its intact function.

Fully automated: DNA-in-Protein (in storage buffer)-out
Purified target protein is eluted in a storage buffer of choice.

Parallel processing system
1-8 different proteins can be synthesized in a single run.

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Procedure and principle

This kit uses oxidative environment during protein synthesis which is different from the conventional reducing environment. The energy source will be continuously provided to the reaction, and the target protein is purified using Ni-NTA magnetic beads.



Via the patented SECF technology, ExiProgen™ synthesizes the target protein.

This kit contains chaperons for helping protein-folding and highly optimized buffer for disulfide bond formation.


Experimental results

1. This kit can synthesize a protein with disulfide bonds resulting in proteins with functional activity.


Left Pannel: Expression and purification of ScFV and rPA.

M: AccuLadder™ Protein Size Marker (Low), 1: ScFV (it has 2 disulfide bonds.), 2: rPA (it has 9 disulfide bonds)
* Amount of purification sample is 1/40 of total sample.

Right panel: rPA synthesized with ExiProgen™ EC2 Protein Synthesis Kit has an enzyme activity.
1: Synthesis sample using ExiProgen™ EC Maxi Protein Synthesis Kit.
2: Synthesis sample using ExiProgen™ EC2 Protein Synthesis Kit.
* rPA activity was measured by incubating with chromogenic substrate S-2288.


 

Application

This kit can be used to produce scFv for antibody screening.


 

Kit contents and storage conditions

This kit consists of two separate boxes, and its components are described in the following table.

Components Storage temperature
Kit 1 Cartridge ① 96 well x 1ea at 4°C
Dialysis tube 1 pack (16 ea/pack)
Disposable filter tip 1 pack (8 ea/pack) at room temperature
Protection cover 1 ea
Kit 2 Cartridge ② 96 well x 1ea at -20°C
(upon receiving,
E. coli extract is
recommended to be
stored at -70°C)
E. coli extract 8-tube strip (Yellow) x 1 ea
Master mix 8-tube strip (Violet) x 1 ea
DEPC DW 8-tube strip (White) x 1 ea
Storage buffer 35 mL bottle x 2 ea
Expression screening set 1 box
Positive control DNA 1.5mL tube x 1 ea


 

 


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