Product name: Recombinant Bovine Enterokinase, His-tag, rbEK-His


Specifications: 200 U (liquid), 1000 U (liquid), 100 μg, 1 mg, special order


Product form: Liquid is 1 U/μl rbEK-His, 50 mM Tris-HCl, 50% glycerol, pH 8.0; the lyophilized powder is 50 mM Tris-HCl (pH 8.0) enzyme solution before lyophilization


Molecular weight: 27.6 kDa (SDS-PAGE detection is about 30 kDa)


Isoelectric point: 5.72


Specific activity: ≥40,000 U/mg, that is, 1 mg of rbEK-His cuts at least 2000 mg of fusion protein.


Purity: ≥95% (SDS-PAGE detection)


A280nm (1 mg/mL): 1.95


Endotoxin: ≤5 EU/mg


Source: Escherichia coli


Storage conditions: -20℃ storage


Validity period: 3 years


Activity definition: 25°C, reaction buffer is 50 mM Tris-HCl (pH 8.0), substrate is 50 μg 1 mg/ml fusion protein with restriction site, which is required for cleavage of 95% of fusion protein within 16 h The amount of enzyme is 1 unit of enzyme activity (1 U).






Related introduction:


Enterokinase (EC 3.4.21.9) is a serine proteolytic enzyme that can efficiently and specifically recognize the Asp-Asp-Asp-Asp-Lys (DDDDK) sequence in the protein, and the C of Lys (Lys, K) The peptide bond is hydrolyzed at the end to produce cleavage, and the trypsinogen is hydrolyzed in the organism and converted into trypsin. Because enterokinase has high specificity and high-efficiency enzyme cleavage characteristics, it is widely used in the development of genetic engineering products.






Natural enterokinase is composed of a 115 kDa heavy chain and a 35 kDa light chain. The heavy chain serves as an anchor to the cell membrane, and the light chain has the complete catalytic activity of the whole enzyme. This product is a fusion protein of the light chain active core region of bovine enterokinase and histidine tag. It is expressed by E. coli secretion technology and has higher specific activity than natural enterokinase. At the same time, it has a histidine tag. After digestion, a nickel column (NRPB07) can be used to remove rbEK-His in the sample, which is especially suitable for digestion of genetic engineering fusion proteins.






Precautions:


1) Some reagents will affect the enzyme activity, such as >2 M Urea, >250 mM NaCl, >20 mM β-ME, >0.1% SDS, >50 mM Imidazole. If the sample contains these components, it needs to be dialyzed to 50 mM Tris-HCl (pH 8.0);


2) Phosphate can inhibit the activity of enterokinase. Avoid using phosphate buffer as the digestion system;


3) Put it on an ice bath after dissolution and try to use it as soon as possible;


4) If you cannot use it up at one time, please prepare the mother liquor before use and divide it into small sizes (to ensure the amount of one test) and store it at -65°C to avoid repeated freezing and thawing affecting protein activity


5) Avoid using strong acids, strong bases, strong oxidants, high-concentration organic solvents and other reagents that are likely to cause protein denaturation;


6) This product is limited to scientific research use only.









M: protein molecular weight standard


Lane 1: rbEK-His

Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance

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