New England Biolabs, P8010S, Factor Xa Protease

Factor Xa cleaves after the arginine residue in its preferred cleavage site Ile-Glu/Asp-Gly-Arg. It will sometimes cleave at other basic residues, depending on the conformation of the protein substrate. The most common secondary site, among those that have been sequenced, is Gly-Arg. Related Categories Proteases Applications Fusion Protein Cleavage,, Target Protein Insolubility ,, Protein Purification, Specification Unit Definition 1 μg of Factor Xa will cleave 50 μg of MBP-fusion protein test substrate, MBP-FXa-Paramyosin-ΔSal to 95% completion in a total reaction volume of 50 μl in 6 hours or less at 25°C in 20 mM Tris-HCl (pH 8.0 @ 25°C) with 100 mM NaCl and 2 mM CaCl2. Usage Concentration 1 mg/ml Removal Factor Xa will bind specifically to benzamidine-agarose. Storage Buffer 50 mM HEPES 500 mM NaCl 2 mM CaCl2 50% Glycerol pH 8 @ 25°C Molecular Weight Theoretical: 43 kDa Deactivation Dansyl-glu-gly-arg-chloromethyl ketone (CALBIOCHEM, #251700) will irreversibly inactivate Factor Xa by covalent attachment at the active site. In a reaction containing 20 μg/ml Factor Xa, 2 μM dansyl-glu-gly-arg-chloromethyl ketone will inactivate >95% of the Factor Xa in 1 minute at room temperature.