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| Place of Origin: | China, Shanghai |
| Brand Name: | YAXINBIO |
| Certification: | NQA ISO 9001:2015 |
| Model Number: | REK08 |
| Minimum Order Quantity: | 100U |
|---|---|
| Price: | 65$/100U |
| Packaging Details: | ice packaging |
| Payment Terms: | T/T |
| Supply Ability: | 10000kU per week |
| Source: | E. Coli | M.W.: | Theoretical MW: 25,850 Da;The Apparent MW On SDS-PAGE: About 27,000 Da |
|---|---|---|---|
| Storage: | Store At -20°C After Delivery. | Specific Activity: | One Unit Is Defined As The Amount Of Enzyme Needed To Cleave 0.5mg Of Fusion Protein In 12 To16 Hours To Get 95% Completion At 25°C In A Buffer 25mMTris-HCl, PH 8.0.Substrate: A Special Fusion Protein. |
| Fusion Protein Concentration: | 0.1-1mg/ml (total Protein Content: 0.5-1.0mg) | Cutting Condition: | Given An Example: 25mM Tris-HCl 8.0 |
| High Light: | enterokinase enzyme,recombinant enzymes |
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Recombinant Enterokinase and Recombinant Bovine Enterokinase
Recombinant Enterokinase
Product Overview
Recombinant Bovine Enterokinase produced in E.coli is a single glycosylated polypeptide chain containing 235 amino acids and having a molecular mass of approximately28 kDa.
Main Features
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M.W. |
25,850 Da |
|
Specific Activity |
One unit is defined as the amount of enzyme needed to cleave 0.5mg of fusion protein in 12 to16 hours to get 95% completion at 25°C in 25mMTris-HCl, pH 8.0. Substrate: a special fusion protein. |
|
Stability |
Keep cool with blue ice during shipping. Remained stable at 25°C for one week without activity lost. No activity lost after 5 cycles of frozen-thawing. |
RECOMMEND USAGE CONDITION:
1) Cutting condition: 25mM Tris-HCl 8.0
Fusion protein concentration: 0.1-1mg/ml (total protein content: 50-100ug)
EK content: 0.1-0.2U
Temperature: 25℃
Time: overnight or 16h-24h for digestion.
If digestion under 2-8°C,prolong the digestion time two folds,or more enzyme added to short the digestion time.
2) 4°C low temperature digestion conditions:
4°C can effectively digest the substrate, but need to extend the digestion time to 48-64h, or increase the amount of 2-3 times the amount of enzyme.
3) Enzyme cleavage optimization and amplification
Optimization: Enzyme cleavage conditions can be optimized, such as buffer pH, fusion protein concentration, EK amount, and digestion time, so that the fusion protein can be digested under stable conditions, SDS-PAGE detection of digestion effect.
Amplification: Select the optimized reaction conditions, according to the conditions such as the proportion of amplification, SDS-PAGE detection of digestion effect.
4) Method for removing recombinant enterokinase: The recombinant enterokinase was removed using trypsin inhibitor affinity chromatography (e.g., sigma T0637).
5) Note: do not recommend cutting at 37°C conditions, there may be non-specific enzyme digestion.
COMMON COMPONENTS INFLUENCE THE ACTION OF ENTEROKINASE
＞200mM imidazole or ＞200mM NaCl or ＞5% glycerin, the reaction may be effected.The following suggestions are given:
1) To receive the optimum result, please dialyze the sample to 25 mMTris-HCl, pH 8.0.
2) If the dialysis is inconvenient, please dilute the sample to ＜100mM imidazole, ＜50mMNaCl, ＜5% glycerin, and the proportion of fusion protein and EK may not be changed (1U:0.5mg fusion protein).
3) If there are one or more components in samples, and cannot be removed, suggest to increase the content of EK in reaction system or extend the reaction time.
Product Information
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Product |
Activity |
Packaging |
Manufacturer |
|
Recombinant Enterokinase |
≥5 u/μl |
100u;500u;1ku;10ku;100ku |
YaxinBio |
Unit Definition
One unit is defined as the amount of enzyme needed to cleave 500 ug of fusion protein in 16 hours to 95% completion at 25°C in a buffer containing 25 mM Tris-HCl, pH 7.6, 50 mM NaCl, and 2 mM CaCl2.
Contact Person: Miss Eland
Tel: +8613482039151
