This form of neutrophil elastase is a salt-free preparation and is supplied in an active form with a specific activity of ≥20 units/mg protein. It is not sterile and has not been tested for cell culture applications. To use it in a cell culture assay, sterile filtration is necessary, which may result in some product retention on the filter.
324681
Elastase, Human Neutrophil
Elastase, Human Neutrophil, CAS 9004-06-2, is a serine protease that degrades elastin, collagen, and proteoglycans
Synonyme(s) :
Elastase, Human Neutrophil
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About This Item
Produits recommandés
Niveau de qualité
Description
Merck USA index - 14, 3535
Forme
lyophilized solid
Activité spécifique
≥20 units/mg protein
Fabricant/nom de marque
Calbiochem®
Conditions de stockage
OK to freeze
Conditions d'expédition
ambient
Température de stockage
−20°C
Description générale
Avertissement
Définition de l'unité
Forme physique
Notes préparatoires
Reconstitution
Remarque sur l'analyse
Autres remarques
Junger, W.G., et al. 1992. Biol. Chem. Hoppe-Seyler 373, 691.
Selak, M.A. 1992. Thromb. Haemost.68, 570.
Baugh, R.J., and Travis, J. 1976. Biochemistry15, 836.
Informations légales
Mention d'avertissement
Danger
Mentions de danger
Conseils de prudence
Classification des risques
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
Organes cibles
Respiratory system
Code de la classe de stockage
11 - Combustible Solids
Classe de danger pour l'eau (WGK)
WGK 3
Point d'éclair (°F)
Not applicable
Point d'éclair (°C)
Not applicable
Certificats d'analyse (COA)
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Les clients ont également consulté
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Hi Is this form of neutrophil elastase active and suitable be used directly in a cell assay? And is it sterile ? Mvh Magdalena
1 answer-
Helpful?
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Has this product been tested for the presence of endotoxin?
1 answer-
Unfortunately, this product has not been tested for the presence for endotoxins
Helpful?
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what is the molecular wight of the enzyme ?
1 answer-
Unfortunately, the molecular weight has not been determined for this product. A number of estimates have been reported in the literature. Please see the link below to a publication that may be helpful:
https://www.sciencedirect.com/science/article/pii/S1016847823175091?via%3DihubHelpful?
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Is this the active form of the enzyme? or does it require activation by cathepsin?
1 answer-
Here are the recommended reconstitution and reaction conditions:
Solubility information:
Reconstitute in 50 mM sodium acetate, pH 5.5, with 200 mM NaCl or in other aqueous buffers. Material is most active near a neutral pH, but to maintain stability upon reconstitution, use in a slightly acidic buffer (pH 5.5) to avoid autolysis.
Following reconstitution, aliquot and freeze (-20°C) for long term storage or refrigerate (4°C) for short-term storage. Acidic stock solutions are stable for up to 1 month at 4°C or for up to 1 year at -20°C.Human Neutrophil Elastase Protocol
1. Dissolve enzyme in reaction buffer: 100 mM Tris-HCl, pH 7.5, 500 mM NaCl.
2. Prepare the substrate stock solution by dissolving Elastase Substrate I (MeOSuc-Ala-Ala-Pro-Val-pNA, Cat. No. 324696) at 50 mM in DMSO.
3. Just prior to assay, dilute the substrate stock solution with an equal volume of 200 mM Tris-HCl, pH 8.0. Mix well.
4. Prepare the assay mixture by combining 960 µl of reaction buffer with 40 µl of diluted substrate stock solution.
5. Add enzyme to assay mixture, mix, and incubate at 25°C.
6. Monitor the change in absorbance at 410 nm. Use an ε of 8800 at 410 nm to calculate the hydrolysis of nitroanilide.Helpful?
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