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T4648

Thrombin aus Rinderplasma

lyophilized powder, 40-500 NIH units/mg protein (biuret)

Synonym(e):

Faktor IIa

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Über diesen Artikel

CAS-Nummer:
UNSPSC Code:
12352204
NACRES:
NA.54
EC Number:
232-648-7
MDL number:
EG-Nummer:
Specific activity:
40-500 NIH units/mg protein (biuret)
Biological source:
bovine plasma
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biological source

bovine plasma

Quality Segment

form

lyophilized powder

specific activity

40-500 NIH units/mg protein (biuret)

mol wt

heavy chain ~33 kDa, light chain ~5 kDa

composition

Protein, 40-60%

color

, white to faint yellow to faint red

UniProt accession no.

shipped in

wet ice

storage temp.

−20°C

Gene Information

cow ... F2(280685)

General description

Thrombin ist die finale Koagulationsprotease bei der Hämostase mit koagulationsfördernder und koagulationshemmender Wirkung.

Application

Thrombin wird für die stellenspezifische Spaltung von rekombinanten Fusionsproteinen mit einer zugänglichen Thrombin-Erkennungsstelle zum Entfernen von Affinitäts-Tags verwendet. Thrombin wurde in einer Studie zur Beurteilung der anhaltenden Hypokoagulabilität bei Patienten mit septischem Schock verwendet.

Biochem/physiol Actions

Serinprotease, die selektiv Arg-Gly-Bindungen in Fibrinogen spaltet, um Fibrin und Fibrinopeptide A und B zu bilden.

Physical form

Lyophilisiertes Pulver mit Natriumchlorid und Tris-HCl, pH-Wert 7,0

Preparation Note

Aktiviert mit Thromboplastin aus Rinderhirn.

Analysis Note

Beim NIH-Assay werden 0,2 ml verdünntes Plasma (1:1 mit Kochsalzlösung) als Substrat und 0,1 ml der Thrombinprobe (stabilisiert in einer 1%igen gepufferten Albuminlösung) auf Basis einer modifizierten Biggs-Methode analysiert. Zur Bestimmung der Thrombinkonzentrationen werden nur Gerinnungszeiten im Bereich von 15 bis 25 Sekunden verwendet.
Die Aktivität wird in NIH-Einheiten ausgedrückt, die aus einem direkten Vergleich mit einem NIH-Thrombin-Referenzstandard (Charge K) gewonnen werden

Other Notes

Die Aktivität wird in NIH-Einheiten ausgedrückt, die sich aus einem direkten Vergleich mit einem NIH-Thrombin-Referenzstandard ergeben.
Für weitere Informationen siehe thrombin at www.sigma-aldrich.com/enzymeexplorer.

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Dieser Artikel
T7326T75131.12374
biological source

bovine plasma

biological source

-

biological source

-

biological source

bovine plasma

specific activity

40-500 NIH units/mg protein (biuret)

specific activity

≥60 NIH units/mg protein (biuret)

specific activity

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

specific activity

50 NIH units/mg dry wt

Gene Information

cow ... F2(280685)

Gene Information

-

Gene Information

cow ... F2(280685)

Gene Information

-

form

lyophilized powder

form

lyophilized powder

form

lyophilized powder

form

solid

shipped in

wet ice

shipped in

-

shipped in

-

shipped in

-

mol wt

heavy chain ~33 kDa, light chain ~5 kDa

mol wt

-

mol wt

heavy chain ~33 kDa, light chain ~5 kDa

mol wt

-


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pictograms

Health hazardExclamation mark

signalword

Danger

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

target_organs

Respiratory system

Lagerklasse

11 - Combustible Solids

flash_point_f

Not applicable

flash_point_c

Not applicable



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Questions

1–10 of 17 Questions  
  1. To make a sterile solution of thrombin, would I first mix the thrombin in BSA (as per the protocol), then filter it using a syringe filter of 0.2um?

    1 answer
    1. Yes, as per the product datasheet, stock solutions can be prepared at a concentration of 100 units/mL in a 0.1% (w/v) BSA solution. Following reconstitution, use a low protein-binding 0.2 um syringe filter to sterilize. Aliquot and store the sterile stock solution in plastic tubes rather than glass to reduce adsorption. Please see the link below to review additional information available in the product information sheet:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/335/094/t4648dat.pdf

      Helpful?

  2. Does this product contain his tag or other tags?

    1 answer
    1. This product is extracted from bovine plasma as described in the datasheet. It is not a recombinant protein and does not contain any tags.

      Helpful?

  3. 10KUは1KUの10倍量でしょうか? 1KUのg数を教えてください。

    1 answer
    1. Yes, the 10KU is 10 times the amount of the 1 KU pack size, the equivalent of 10,000 units. The gram quantity is lot-specific and dependent upon the protein content and the unit activity per milligram protein. This product may range from 40 - 60% protein and the activity may range from 40 - 300 units per milligram protein. At the minimum of 40% protein and 40 units per milligram protein, the amount of material per 10KU bottle is 625 mg. Please see the link below to review a sample or lot-specific Certificate:
      https://www.sigmaaldrich.com/product/sigma/t46483product-documentation

      Helpful?

  4. Is T4648 thrombin sterile?

    1 answer
    1. This product is not a sterile powder. Prepared solutions may be filter-sterilized.

      Helpful?

  5. Is this product (T4648-1KU) suitable for cell culture experiments? endotoxin free?

    1 answer
    1. 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.

      Helpful?

  6. 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
    1. 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.

      Helpful?

  7. How do i dissolve the lyophilized powder of T4648 bovine thrombin of 40-300 NIH units/mg

    1 answer
    1. 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.pdf

      Helpful?

  8. What is the bottle information for T4648-1KU and T4648-10KU?

    1 answer
    1. Both products are packaged in a 20 ML Amber glass bottle with a 24-400 thread.

      Helpful?

  9. I was cleave gfp from my protein which is dissovled in PBS. Should I make thrombin stock in pbs or bsa?

    1 answer
    1. 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.pdf

      Helpful?

  10. What is the purity of this product?

    1 answer
    1. 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?

1–10 of 17 Questions  

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