Factor Xa Protease |
Catalog No.GC26287 |
For Factor Xa, fusion protein cleavage is carried out at a w/w ratio of 1% the amount of fusion protein (e.g., 1 mg Factor Xa for a reaction containing 100 mg fusion protein).
Products are for research use only. Not for human use. We do not sell to patients.
Sample solution is provided at 25 µL, 10mM.
For Factor Xa, fusion protein cleavage is carried out at a w/w ratio of 1% the amount of fusion protein (e.g., 1 mg Factor Xa for a reaction containing 100 mg fusion protein). The reaction mixture can be incubated for 3 hours to several days, at room temperature or 4°C. Depending on the particular fusion protein, the amount of protease can be adjusted within the range of 0.1–5.0%, to get an acceptable rate of cleavage. Factor Xa will cleave at non-canonical sites in some proteins; for some fusions, there is a correlation between instability of the protein of interest in and cleavage at additional sites (unpublished observations). Presumably this cleavage activity depends on the three dimensional conformation of the fusion protein. For fusions that are resistant to cleavage, two strategies can sometimes help. Inclusion of small amounts of SDS (0.005–0.05%) in the reaction appears to relax the fusion enough to allow for cleavage in some cases. The window of SDS concentrations that work can be small, so a pilot titration with different SDS concentrations is necessary. Another strategy that sometimes helps is to denature the fusion to render the protease site accessible to cleavage.
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