Preparation of the stock solution in Tris-Nacl buffer at pH 7.5 has not been tested and requires validation through literature review. The amount of thrombin required for protein cleavage depends on the specific protein and expression system and should be optimized accordingly. As a starting point, use 1 to 10 units of thrombin per milligram of recombinant protein.
T7326
Thrombin from bovine plasma
≥60 NIH units/mg protein (biuret)
Synonym(s):
Factor IIa
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About This Item
CAS Number:
UNSPSC Code:
12352204
NACRES:
NA.54
MDL number:
Specific activity:
≥60 NIH units/mg protein (biuret)
Pricing and availability is not currently available.
form
lyophilized powder
Quality Segment
specific activity
≥60 NIH units/mg protein (biuret)
composition
Protein, ≥50%
storage temp.
−20°C
General description
Thrombin is the final coagulation protease in regard to hemostasis, promoting both procoagulant and anticoagulant effects. Thrombin comprises of A-chain and B-chain with the catalytic pocket. The C-terminal regions has a fibrinogen recognition site (FRS) and a heparin binding site (HBS). The main function of thrombin is cleavage of fibrinogen to fibrin. It coordinates the coagulation cascade and coordinates with protease-activated receptors (PARs) to regulate physiological functions. High levels of thrombin causes neurotoxicity in dopaminergic neurons and contribute to the progression of parkinsons disease. Altered thrombin levels modulates coagulation pathway in multiple sclerosis. Patients with coronary artery disease (CAD) show elevated levels of thrombin.
Application
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 evaluate coagulation abnormalities in acute liver failure. [1]
Thrombin from bovine plasma may be used:
as revitalization model to mimic blood fabrication in root canal
- for the generation of fibrin from fibrinogen embedded in microbubbles
- for the cleavage of hydrogenase maturation protein (HypF-N) from the nickel resin
as revitalization model to mimic blood fabrication in root canal
Biochem/physiol Actions
Serine protease that selectively cleaves Arg-Gly bonds in fibrinogen to form fibrin and fibrinopeptides A and B.
Physical form
Lyophilized powder containing sodium chloride and Tris-HCl, pH 7.0
Preparation Note
Traditional thrombin products are activated with bovine brain thromboplastin, whereas T7326 is activated with bovine lung thromboplastin and does not contain any bovine brain products in its preparation.
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
Activity is expressed in NIH units obtained by direct comparison to a NIH thrombin reference standard.
View more information on thrombin at www.sigma-aldrich.com/enzymeexplorer.
1 of 1
This Item | |||
|---|---|---|---|
| specific activity ≥60 NIH units/mg protein (biuret) | specific activity ≥2800 NIH units/mg protein (E1%/280, 18.3) | specific activity 400-1000 NIH units/mg protein | specific activity 1500-3500 NIH units/mg protein (E1%/280, 18.3) |
| form lyophilized powder | form lyophilized powder | form lyophilized powder | form lyophilized powder |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
| composition Protein, ≥50% | composition - | composition - | composition - |
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Storage Class
11 - Combustible Solids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, type N95 (US)
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Can the stock be prepared in Tris-Nacl buffer of pH 7.5? And in what ratio of protein should cleavage and thrombin be used, and what is the ideal condition for cleavage?
1 answer-
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Dear team I would like to ask about this product T7326 (Thrombin from bovine plasma ) ,1000U. What is the best way of preparation? If I would like to prepare 2.5U/L How should I do that? Regards
1 answer-
Stock solutions can be prepared at a concentration of 100 units/mL in a 0.1% (w/v) BSA solution. Further dilutions can be prepared fresh to the final desired working concentration.
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