This product is not tested for endotoxin content. However, it has been cited for cell culture application in the publication: Neuroreport. 2015 May 6;26(7):416-23.
T4648
Trombina from bovine plasma
lyophilized powder, 40-500 NIH units/mg protein (biuret)
Sinónimos:
Factor IIa
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About This Item
En este momento no podemos mostrarle ni los precios ni la disponibilidad
Productos recomendados
origen biológico
bovine plasma
Nivel de calidad
Formulario
lyophilized powder
actividad específica
40-500 NIH units/mg protein (biuret)
mol peso
heavy chain ~33 kDa
light chain ~5 kDa
composición
Protein, 40-60%
Nº de acceso UniProt
Condiciones de envío
wet ice
temp. de almacenamiento
−20°C
Información sobre el gen
cow ... F2(280685)
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Descripción general
La trombina es la proteasa final de la coagulación con respecto a la hemostasia, promoviendo tanto efectos procoagulantes como anticoagulantes.
Aplicación
La trombina se utiliza para la rotura específica de sitio de las proteínas de fusión recombinantes que contienen un sitio de reconocimiento de trombina accesible para eliminar las etiquetas de afinidad. La trombina se ha utilizado en un estudio para evaluar la hipocoagulabilidad persistente en pacientes con choque séptico. [1]
Acciones bioquímicas o fisiológicas
Serín proteasa que hidroliza selectivamente los enlaces Arg-Gly en el fibrinógeno para formar fibrina y fibrinopéptidos A y B.
Definición de unidad
La actividad se expresa en unidades NIH obtenidas por comparación directa con un patrón de referencia de la trombina NIH.
Forma física
Polvo liofilizado que contiene cloruro de sodio y Tris-HCl, pH 7,0
Nota de preparación
Se activa con tromboplastina cerebral bovina.
Nota de análisis
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.
Otras notas
Véase más información sobre la trombina en www.sigma-aldrich.com/enzymeexplorer.
Inhibidor
Referencia del producto
Descripción
Precios
Palabra de señalización
Danger
Frases de peligro
Consejos de prudencia
Clasificaciones de peligro
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
Órganos de actuación
Respiratory system
Código de clase de almacenamiento
11 - Combustible Solids
Clase de riesgo para el agua (WGK)
WGK 2
Punto de inflamabilidad (°F)
Not applicable
Punto de inflamabilidad (°C)
Not applicable
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Is this product (T4648-1KU) suitable for cell culture experiments? endotoxin free?
1 answer-
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I just bought this thrombin to cleave HisTag from recombinant proteins but I am struggling in understanding how much /mg of purified recombinant protein I have to use. Do you have any suggestion? Thanks a lot
1 answer-
The amount of thrombin to use depends on the protein expression system. Optimization is needed for each specific protein, and 1 to 10 units per mg of recombinant protein can be used as a starting point.
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How do i dissolve the lyophilized powder of T4648 bovine thrombin of 40-300 NIH units/mg
1 answer-
This product is soluble in water at 10 mg/mL. Stock solutions may be prepared at 100 units/mL in water containing 0.01% BSA. Please see the link below to review the product datasheet for additional information, including solution stability and storage:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/335/094/t4648dat.pdfHelpful?
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What is the bottle information for T4648-1KU and T4648-10KU?
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Both products are packaged in a 20 ML Amber glass bottle with a 24-400 thread.
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I was cleave gfp from my protein which is dissovled in PBS. Should I make thrombin stock in pbs or bsa?
1 answer-
The recommended preparation instructions, per the product information sheet, are 10 mg/mL in water or 100 units/mL in a 0.01% BSA solution. Stock solutions of either aforementioned recommendations can then be added to PBS at the desired usage concentration. The addition of BSA to the stock solution will aid in long-term product stability when stored at -20 deg. C. See the link below to review the product information sheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/335/094/t4648dat.pdfHelpful?
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What is the purity of this product?
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The purity of this product is not determined. The biuret method is used to verify the protein content. In general, products with higher activity levels can be considered higher in 'purity'. See the link below to other bovine thrombin products for comparison:
https://www.sigmaaldrich.com/substance/thrombinfrombovineplasma123459002044.Helpful?
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What is the isoelectric point of Product T4648, Thrombin from bovine plasma?
1 answer-
The isoelectric point of thrombin from bovine plasma is 7.05.
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What is the solubility of Product T4648, Thrombin from bovine plasma?
1 answer-
Thrombin from bovine plasma is soluble in water at 10 mg/mL.
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What is the solution stability of Product T4648, Thrombin from bovine plasma?
1 answer-
When stock solutions of thrombin from bovine plasma are prepared at a concentration of 100 units/mL in 0.1% (w/v) BSA, they are stable for one week at 0-5°C. Since thrombin solutions adsorb to glass, it is recommended to aliquot the solutions in plastic tubes and store at -20°C for long term storage.
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What is the cross reactivity of Thrombin?
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Thrombin from any vertebrate species will clot the fibrinogen of virtually any other species. Fibrinogen from any mammalian source will cross-react with thrombin from any mammalian source, but it is not well known how thrombin and fibrinogen from different animals differ in their sequence. When any mammalian thrombin is injected into a different mammal, clotting will occur.
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