The Enterokinase Cleavage Capture Kit is designed for highly specific cleavage of fusion proteins followed by rapid, affinity-based capture and removal of enterokinase. Following cleavage of the target protein, rEK is removed with >99% efficiency from the reaction by affinity capture on EKapture™ Agarose. Following capture of rEK, the EKapture Agarose is removed by spin filtration, because the same buffer conditions are used for both cleavage and capture no buffer changes are necessary.
A Cleavage Control Protein is included in each kit for conducting control digests in parallel with experimental samples, or to test cleavage under customized buffer conditions. rEK cleaves the 48-kDa Cleavage Control Protein into two proteolytic fragments of 32 kDa and 16 kDa, each of which are easily visualised by standard SDS-PAGE followed by Coomassie blue staining. The Cleavage Control Protein also features an amino terminal S•Tag™ sequence enabling sensitive detection of the 16 kDa proteolytic product with Western blot reagents.
Delivery information: Kit contains 50 U recombinant Enterokinase (in 200 mM NaCl, 20 mM Tris-HCl, 2 mM CaCl₂, 50% glycerol, pH 7,4); 10 µg Cleavage Control Protein (in 200 mM NaCl, 20 mM Tris-HCl, 20 mM EDTA, 50% glycerol, pH 7,5); 2 ml 1X rEK Dilution/Storage Buffer (200 mM NaCl, 20 mM Tris-HCl, 2 mM CaCl₂, 50% glycerol, pH 7,4); 5 ml 10X rEK Cleavage/Capture Buffer (500 mM NaCl, 200 mM Tris-HCl, 20 mM CaCl₂, pH 7,4); 1,5 ml EKapture™ Agarose (3 ml 50% slurry in phosphate buffer, 500 mM NaCl, 0,02% Sodium Azide, pH 7,3); and 10 spin filters (2 ml capacity).
