T7201

Thrombin from bovine plasma

lyophilized powder, ≥2,000 NIH units/mg protein (E1%/280 = 19.5)

• Synonym(s): Factor IIa
• CAS Number: 9002-04-4
• Enzyme Commission number: 3.4.21.5 (BRENDA, IUBMB)
• MDL number: MFCD00166053
• UNSPSC Code: 12352204
• NACRES: NA.54

Properties

• form: lyophilized powder
• specific activity: ≥2,000 NIH units/mg protein (E1%/280 = 19.5)
• mol wt: heavy chain ~31 kDa; light chain ~6 kDa
• storage temp.: −20°C

Description

General description

Thrombin is the final coagulation protease in regard to hemostasis, promoting both procoagulant and anticoagulant effects. Thrombin consists of A-chain and a catalytic B-chain with the catalytic pocket comprising of His57, Asp102, Ser195 and Ser214. It has a fibrinogen recognition site (FRS) and a heparin binding site (HBS) in the C-terminus. The main function of thrombin is cleavage of fibrinogen to fibrin. It coordinates the coagulation cascade and binds to diverse substrates. It coordinates with protease-activated receptors (PARs) to accomplish the physiological functions. High levels of thrombin elicit neurotoxicity in dopaminergic neurons and contribute to the progression of Parkinson′s disease. Altered thrombin levels modulates coagulation pathway in multiple sclerosis. Patients with coronary artery disease (CAD) show elevated levels of thrombin.

Application

Thrombin from bovine plasma has been used in the cleavage of the maltose binding protein tag in mini-calpain-5 fusion protein and in thrombin/hirudin spiking experiments in serum samples.

Thrombin is used for site specific cleavage of recombinant fusion proteins containing an accessible thrombin recognition site for removal of affinity tags. Thrombin has been used in a study to assess a novel antiplatelet therapy in acute coronary syndromes.

Biochem/physiol Actions

Serine protease that selectively cleaves Arg-Gly bonds in fibrinogen to form fibrin and fibrinopeptides A and B.

Unit Definition

Activity is expressed in NIH units obtained by direct comparison to a NIH thrombin reference standard.

Physical form

Lyophilized from saline sodium citrate buffer, pH 6.5

Preparation Note

Traditional thrombin products are activated with bovine brain, whereas T7201 is activated with bovine lung and does not contain any bovine brain products.

Analysis Note

Activity is expressed in NIH units obtained by direct comparison to a NIH Thrombin Reference Standard, Lot K.

The NIH assay procedure uses 0.2 mL of diluted plasma (1:1 with saline) as a substrate and 0.1 mL of thrombin sample (stabilized in a 1% buffered albumin solution) based on a modification of the method of Biggs. Only clotting times in the range of 15-25 seconds are used for determining thrombin concentrations.

Other Notes

View more information on thrombin at www.sigma-aldrich.com/enzymeexplorer.

Safety Information

• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 2
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable
• Personal Protective Equipment: dust mask type N95 (US), Eyeshields, Gloves