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Recombinant Enterokinase


HBP009003-1 2KU
HBP009003-2 20KU
HBP009003-3 200KU
HBP009003-4 2MU
HBP009003-5 20MU


Cat. No: HBP009003

Storage stability: stored at -25°C~-15°C, stable within 2 years.  

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Product description

Enterokinase (EK) is a highly specific serine protease that specifically recognizes the Asp- Asp-Asp-Asp-Lys sequence and cleavages at the C-terminus of Lys. Hzymes recombinant enterokinase (rEK) (containing 6×His-tag) is expressed by Escherichia coli and especially suitable for the production of peptide drugs, such as GLP-1 and its analogues.


Product application

1. Used to remove the N-terminal fusion tag of the recombinant protein.

2. For the activation of trypsinogen.

3. Production of recombinant pharmaceutical.

proteins. Such as BNP, GLP-1 and its analogues.


Transportation and storage

Storage stability: stored at -25°C~-15°C, stable within 2 years.

Transport stability: Transportation of Dry Ice.


Unit definition

1U is defined as the amount of enzyme that will cleave 95% the fusion protein (0. 5mg) in 12-16h at 25ºC , in a reaction buffer containing 25 mM Tris-HCl (pH 8. 0).


Quality control

• Appearance: clear, colorless to light yellow liquid.

• Purity (SDS-PAGE): ≥95%.

• Molecular weight: ~27. 0 kDa.


Usage

•  Buffer: 25 mM Tris-HCl (pH 8. 0).

•  Enzyme Digestion Ratio: 0. 1-1 U can be used to cleave fusion proteins: 0. 1-1 mg.

•  Reaction Time: Incubate at 25 C for 12-16h.

•  Incubation at 4 C requires a longer time or an increase in the dosage of enzyme.

•  Reaction System: The digestion system can be optimized, such as ionic strength, pH, protein concentration, enterokinase dosage, digestion temperature, digestion time, etc.

•  Digestion Verification: Western Blot, SDS- PAGE, RP-HPLC, etc.

•  Enterokinase Removal: His-tag enterokinase.


can be removed by Ni column affinity chromatography adsorption, and an ion- exchange resin binds to DEAE at pH 8. 0.



Notes

•  The enzymatic digestion is affected under conditions of > 200 mM imidazole, > 200 mM NaCl, or > 5% glycerol.

•  In imidazole < 100 mM, NaCl < 50 mM, glycerol < 5%, the amount of enzyme and the ratio of protein unchanged.

•  If the system contains other components that affect the digestion efficiency, they can be removed by dialysis, or diluted to reduce the inhibitor concentration, or increased the amount of enzyme or prolonged the digestion time.