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.
价格与库存信息目前不能提供
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质量水平
描述
Merck USA index - 14, 3535
表单
lyophilized solid
比活
≥20 units/mg protein
制造商/商品名称
Calbiochem®
储存条件
OK to freeze
运输
ambient
储存温度
−20°C
一般描述
弹性蛋白酶，人中性粒细胞，CAS 9004-06-2，是一种降解弹性蛋白、胶原蛋白和蛋白多糖的丝氨酸蛋白酶
来自人中性粒细胞的天然弹性蛋白酶。一种降解弹性蛋白、胶原蛋白和蛋白聚糖的丝氨酸蛋白酶。它的过度表达与肺气肿，类风湿关节炎和囊性纤维化的发展有关。
来自人中性粒细胞的天然弹性蛋白酶。降解胶原蛋白、弹性蛋白和蛋白聚糖。弹性蛋白酶与肺气肿和类风湿关节炎的发展有关。
警告
毒性：标准处理（A）
单位定义
一个单位定义为在25°C，pH 8.0下，每分钟水解1.0 µmol MeO-Suc-Ala-Ala-Pro-Val-pNA（目录号454454）的酶量。
外形
无盐。
制备说明
由已经认证检测显示对HBsAg以及HIV和HCV抗体呈阴性的血液而制备。
重悬
复溶后，等分并冷冻长期保存（-20°C），或冷藏（4°C）短期保存。酸性储备溶液在4°C下可稳定保存长达1个月，在-20°C下可稳定保存长达1年。
用50 mM醋酸钠（pH 5.5）和200 mM NaCl或其他水性缓冲液复溶。材料在中性pH值附近的活性最高，但为了在复溶时保持稳定性，请在弱酸性缓冲液（pH 5.5）中使用以避免自溶。
分析说明
通过SDS-PAGE获得主条带
其他说明
Siedle, B., et al. 2002.Biorg.Med. Chem.10, 2855.
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.
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.
法律信息
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
警示用语：
Danger
危险分类
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
靶器官
Respiratory system
储存分类代码
11 - Combustible Solids
WGK
WGK 3
闪点(°F)
Not applicable
闪点(°C)
Not applicable
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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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