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
T4648
Thrombine from bovine plasma
lyophilized powder, 40-500 NIH units/mg protein (biuret)
Synonyme(s) :
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A propos de cet article
Numéro CAS:
UNSPSC Code:
12352204
NACRES:
NA.54
EC Number:
232-648-7
MDL number:
Specific activity:
40-500 NIH units/mg protein (biuret)
Biological source:
bovine plasma
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Service technique
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Laissez-nous vous aiderbiological 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
La thrombine est la dernière protéase dans la cascade de coagulation pour l′hémostase ; elle présente des effets à la fois procoagulants et anticoagulants.
Application
La thrombine est utilisée pour le clivage de protéines de fusion recombinantes contenant un site de reconnaissance correspondant accessible - le clivage spécifique au site permet d′éliminer les étiquettes d′affinité. La thrombine a été utilisée dans une étude pour évaluer l′hypocoagulabilité persistante chez les patients en choc septique.
Biochem/physiol Actions
Sérine protéase qui clive sélectivement les liaisons Arg-Gly du fibrinogène pour former la fibrine et les fibrinopeptides A et B.
Physical form
La poudre lyophilisée contient du chlorure de sodium et du TRIS HCl, pH 7,0.
Preparation Note
Activée par la thromboplastine de cerveau bovine.
Analysis Note
La procédure d′essai selon NIH utilise 0,2 ml de plasma dilué (1:1 dans une solution saline) comme substrat et 0,1 ml d′échantillon de thrombine (stabilisé dans une solution tamponnée contenant 1 % d′albumine) et est une modification de la méthode de Biggs. Seuls les temps de coagulation dans la plage de 15 à 25 secondes sont utilisés pour déterminer les concentrations en thrombine.
L′activité est exprimée en unités de NIH et obtenue par comparaison directe à une norme de référence de thrombine NIH, Lot K.
Other Notes
L′activité est exprimée en unités de NIH et obtenue par comparaison directe à une norme de référence de thrombine NIH.
Pour en savoir plus sur la thrombine, rendez-vous sur www.sigma-aldrich.com/enzymeexplorer.
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Cet article | |||
|---|---|---|---|
| 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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signalword
Danger
hcodes
Hazard Classifications
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
target_organs
Respiratory system
Classe de stockage
11 - Combustible Solids
flash_point_f
Not applicable
flash_point_c
Not applicable
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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-
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Does this product contain his tag or other tags?
1 answer-
This product is extracted from bovine plasma as described in the datasheet. It is not a recombinant protein and does not contain any tags.
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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-
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-documentationHelpful?
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Is T4648 thrombin sterile?
1 answer-
This product is not a sterile powder. Prepared solutions may be filter-sterilized.
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Is this product (T4648-1KU) suitable for cell culture experiments? endotoxin free?
1 answer-
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.
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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?
1 answer-
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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