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Disulfide Bond PLUS Expression Kit | CellFree Sciences

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Intended to make proteins having disulfide bonds

Disulfide Bond PLUS Expression Kit

Entry kit, designed for ease use to test reaction conditions for disulfide bond formation

Disulfide Bond PLUS Expression Kit 2 en

Disulfide Bond PLUS Expression Kit

This product provides special reaction conditions to better support the formation of disulfide bonds in proteins expressed in the wheat germ cell-free expression system. Reaction conditions have been revised by adding a human endoplasmic reticulum oxidoreductase (Ero1α) and a Protein disulfide isomerase (PDI) to the translation reaction while using a different buffer composition for bilayer formation. Learn more here on how to we support disulfide bond formation in our expression system.

 

Before using this product, confirm whether your target protein requires disulfide bond formation for correct folding and activity. Do not use this product when working with the GST-tag for later purification. Disulfide bonds are not stable when using glutathione containing buffers during the elution step.

Features:

  • Reagents necessary for each reaction are already premixed (Ero1a and PDI mixture provided separately)

  • Synthesizing proteins with just a few pipetting steps

  • Target proteins can be confirmed through SDS-PAGE/CBB staining

  • Kit comes with expression vector, and primers to support template preparation
  • Kit provided positive control to confirm formation of disulfide bonds

Recommended for:

  • Users who want to prepare proteins having disulfide bonds

  • Users who have difficulties with a protein of interest can try these reaction conditions when no prior information on disulfide bond formation is available to test better solubility and activity
  • Users who want to test reaction conditions for scaling up protein expression

 

The basic reagents necessary for each reaction step are pre-packaged into tubes, so there is no need to prepare any reagents.

Enzyme mix is provided separately.

 

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Flow of operations

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Specifications

Product Number

Product Name

Volume

CFS-EDX-DB

Disulfide Bond PLUS Expression Kit

230 µl × 8 reactions

 

 

Product Composition

Reagent Name

Concentration

Volume

Quantity

pEU-E01-MCS

1 µg/µL

5 µL

1

pEU-E01-His-tPA(297-562)(Positive Control) 1 µg/µL 5 µL 1
PCR primer SPU 1 µM 100 µL 1
PCR primer deSP6E01 10 nM 100 µL 1
WEPRO for DB PLUS *1 - 16 µL 8
SUB-AMIX SGC DTT-free 1x 200 µL 8
Transcription Premix LM *2 1x 18 µL 8
PDI & Ero1α mix - 40 µL 1
Aluminum seal - - 2
Storage Condition

 -80 ℃

 

*1 : WEPRO for DB PLUS is based on WEPRO7240H, with the addition of Creatine Kinase and SUB-AMIX SGC DTT-free.

*2 : Transcription Premix LM is based on Transcription Buffer LM, with the addition of NTP Mix, SP6 RNA Polymerase and RNase Inhibitor.

Product-related Information

All our products are for research purpose only; not for use in diagnostic testing and use in human.

Contact us for more information and help on the use of our products.

 

Applicasion Notes

Synthesis of tissue plasminogen activator (tPA) protease domain with retained activity

 

Synthesis of IgG Antibody Fab fragment

 

Related Products

WEPRO7240H Core Kit 2

WEPRO7240H Core Kit

Required for use of  Disulfide Bond Enhancer Enzyme Set to prepare proteins with disulfide bonds at different reaction scales.

Disulfide Bond Enhancer Enzyme Set2 en

Disulfide Bond Enhancer Enzyme Set

Extension kit to be used in combination with WEPRO7240H Core Kit to support expression of proteins having disulfide bonds.

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