Recombinant enterokinase Molecular weight: 25,850DA (theoretical MW) Features:High purityThis product is a high purity recombinant bovine enterokinase. The enzyme was purified by HPLC with high purity.
High specificity1) Specific proteases cleave the carboxy-terminal site of lysine with four aspartic acids in front: aspartic acid - aspartic acid - aspartic acid - aspartic acid - lysine (DDDDK)2) No other proteases, non-specific cuttingImage 1: SDS-PAGE results of purified enterokinase with a single main bandFigure 2. SDS-PAGE analysis of 50µg substrate treated with EK of 0.1U for 0min,10 min,20 min,30 min,40 min,50 min,60 minEnzyme digestion conditions:According to the definition of enzyme activity, an example of recombinant EK digestion conditions was given in 25mM Tris-HCl 8.0 systemItem valueActivity (defined by 1U) : The amount of enzyme required for 95% cleavage of 0.50mg fusion protein in 25mM Tris-HCl pH8.0 buffer at 25℃ for 12h to 16h.Fusion protein concentration 0.1-1mg/ml (total protein 0.50-1.0mg)The amount of recombinant EK is 1.0-2.0UEnzyme digestion temperature 25℃Enzyme digestion time overnight enzyme digestion, or complete enzyme digestion needs 12h-16hCommon factors affecting enterokinase activity:At >200mM imidazole, or >200mM NaCl, or >5% glycerol, the enzyme digestion effect was affected. The following recommended methods can be used for enzyme digestion:To obtain ideal digestion results, the samples were dialyzed into 25mM Tris-HCl pH8.0 buffer before digestion.If dialysis is inconvenient, the sample can be diluted, imidazole content is less than 100mM, NaCl concentration is less than 50mM, glycerol concentration is less than 5%, enzyme digestion, the amount of enzyme and protein ratio remains unchanged (i.e., 1U enzyme digestion 0.5mg protein).If the sample solution contains one or more of the above ingredients, and it is inconvenient to remove, appropriate increase of enzyme amount or extension of enzyme digestion time can also achieve better enzyme digestion effect.Stability:It can be transported at room temperature and stored stably for a week at 37℃.The enzyme liquid can be frozen and thawed several times, which has no effect on enzyme activity.Source and Use:This product is derived from recombinant Escherichia coli and is used to remove fusion proteins located at the N-terminal of proteins to remove unwanted fusion tags.Storage conditions:-20℃, avoid light moisture-proof sealed dry.
Conditions of transportation:2~8℃ transport 

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