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T7201

Sigma-Aldrich

Thrombin from bovine plasma

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

Synonym(s):

Factor IIa

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About This Item

CAS Number:
Enzyme Commission number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54

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

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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.

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

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Small-angle X-ray scattering of calpain-5 reveals a highly open conformation among calpains
Gakhar L, et al.
Journal of Structural Biology, 196(3), 309-318 (2016)
Thrombin-mediated degradation of parathyroid hormone in serum tubes
Laulu SL, et al.
Clinica Chimica Acta; International Journal of Clinical Chemistry, 437, 191-196 (2014)
Karin A L Müller et al.
Current vascular pharmacology, 10(5), 606-618 (2012-06-26)
Acute coronary syndromes (ACS) are triggered by enhanced platelet activation and aggregation. Hence, a cornerstone of successful secondary prevention in ACS is an effective platelet inhibition. Additionally, coronary interventions (PCI) lead to even increased artherothrombotic risks, another challenge in preventing
Lokesh Gakhar et al.
Journal of structural biology, 196(3), 309-318 (2016-07-31)
Calpain-5 is a calcium-activated protease expressed in the retina. Mutations in calpain-5 cause autosomal dominant neovascular inflammatory vitreoretinopathy (ADNIV, OMIM#193235). The structure of calpain-5 has not been determined, thus hindering the investigation of its proteolytic targets and pathological role in
Thrombin generation correlates with disease duration in multiple sclerosis (MS): Novel insights into the MS-associated prothrombotic state
Parsons MEM, et al.
Multiple Sclerosis Journal, 3(4), 84-84 (2017)

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