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
Trombina from bovine plasma
≥60 NIH units/mg protein (biuret)
Sinónimos:
Factor IIa
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Número CAS:
UNSPSC Code:
12352204
NACRES:
NA.54
MDL number:
Specific activity:
≥60 NIH units/mg protein (biuret)
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Servicio técnico
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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.
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
Serín proteasa que hidroliza selectivamente los enlaces Arg-Gly en el fibrinógeno para formar fibrina y fibrinopéptidos A y 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
En el procedimiento de análisis NIH se utilizan 0,2 ml de plasma diluido (1:1 con disolución salina) como sustrato y 0,1 ml de muestra de trombina (estabilizada en una disolución de albúmina tamponada al 1 %), basado en una modificación del método de Biggs. Para determinar las concentraciones de trombina sólo se utilizan los tiempos de coagulación comprendidos entre 15 y 25 segundos.
La actividad se expresa en unidades NIH obtenidas por comparación directa con el patrón de referencia de la trombina NIH, lote K.
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.
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Este artículo | |||
|---|---|---|---|
| description ≥60 NIH units/mg protein (biuret) | description lyophilized powder, 40-500 NIH units/mg protein (biuret) | description lyophilized powder, 600-2,000 NIH units/mg protein (biuret) | description lyophilized powder, ≥2,000 NIH units/mg protein (E1%/280 = 19.5) |
| specific activity ≥60 NIH units/mg protein (biuret) | specific activity 40-500 NIH units/mg protein (biuret) | specific activity 600-2,000 NIH units/mg protein (biuret) | specific activity ≥2,000 NIH units/mg protein (E1%/280 = 19.5) |
| 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 300 | Quality Level 300 | Quality Level 200 |
| composition Protein, ≥50% | composition Protein, 40-60% | composition - | composition - |
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Clase de almacenamiento
11 - Combustible Solids
wgk
WGK 3
flash_point_f
Not applicable
ppe
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flash_point_c
Not applicable
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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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