What is factor Xa protease?
Factor Xa Protease is a site-specific endoprotease that preferentially cleaves the C-terminal peptide bond of the recognition sequence Ile-Glu-Gly-Arg. Factor Xa Protease consists of two polypeptides linked by a disulfide bond.
What does Factor Xa cleave?
Factor Xa is a serine protease which cleaves prothrombin to generate thrombin and lies at the crossroads of the extrinsic and intrinsic coagulation pathway.
Is Xa a protease?
Product Source. Factor Xa Protease is purified from bovine plasma and activated by treatment with the activating enzyme from Russell’s viper venom.
How do I get rid of factor Xa?
Thrombin or Factor Xa is removed by connecting a second (Benzamidine) column. Tagged proteases are removed on the same column as used for capture. An affinity tag can be cleaved from your target protein by first eluting the protein from an affinity resin and then adding a protease.
What is the factor Xa site for?
The factor Xa protease can be used in biochemistry to cleave off protein tags that improve expression or purification of a protein of interest. Its preferred cleavage site (after the arginine in the sequence Ile-Glu/Asp-Gly-Arg, IEGR or IDGR) can easily be engineered between a tag sequence and the protein of interest.
What is factor Xa site?
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.
How do you remove his tag from protein?
His-tag removal from protein using TEV Protease
- Dialyze the protein against 20 mM Tris-HCl, pH 7.5.
- Determine the protein concentration.
- Combine 15 μg of protein and H2O (if necessary) to make a 45 μl total reaction volume.
- Add 5 μl of TEV Protease Reaction Buffer (10X) to make a 50 μl total reaction volume.
How do you get rid of protease?
TEV Protease contains a polyhistidine tag at its N-terminus and can be removed from the reaction by immobilized metal affinity chromatography, such as NEBExpress Ni-NTA Magnetic Beads (NEB #S1423), NEBExpress Ni Spin Columns (NEB #S1427), or NEBExpress Ni Resin (NEB #S1428).
What do factor Xa inhibitors do?
Factor Xa inhibitors are a type of anticoagulant (blood thinning drugs) that work by binding selectively and reversibly to the clotting factor Xa. Factor Xa plays a crucial role in the blood clotting mechanism when you get an injury by forming a mesh to prevent loss of blood.
What is the active enzyme factor Xa?
Factor X is the zymogen of a serine protease. The activated enzyme (factor Xa) is an Arg-specific serine protease related to trypsin, but has much narrower substrate specificity.
What is the purpose of a His-tag?
His Tags allow researchers to selectively extract a protein of interest from the thousands of other proteins found in either a cell or cell lysate.
Why are protease inhibitors necessary?
Protease inhibitors are a type of antiretroviral therapy (ART) medication. These medications reduce the amount of HIV in a person’s blood. This helps control the virus and maintain a person’s overall health.