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产品中心 药物靶点和Fc受体 药物靶点

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MMP-9 His Tag Protein, Mouse

MMP9,CLG4B,GELB,MANDP2,Gelatinase B

价格 ￥1,600.00 （ 供应商现货 ： 3-5个工作日）
货号 UA016087
规格
数量
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产品介绍

产品介绍 评论(0)

产品规格

  • 物种

    Mouse

  • 分子别名

    MMP9, CLG4B, GELB, MANDP2, Gelatinase B

  • Accession

    P41245

  • 表达序列

    Ala20-Pro730 with His Tag at C-Terminus

  • 表达宿主

    HEK293

  • 分子量

    100-130kDa (Reducing)

  • 纯度

    > 95% by SDS-PAGE,>90% by HPLC

  • 内毒素含量

    <0.1EU/μg

  • 活性

    Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2. The specific activity is >3,000 pmol/min/µg, as measured under the described conditions.

  • 标记

    Unconjugated

  • 标签

    His Tag

  • 性状

    Lyophilized Powder

  • 缓冲体系

    PBS, pH7.4, 5% trehalose

  • 溶解方法

    Reconstitute at 0.1-1 mg/ml according to the size in ultrapure water after rapid centrifugation.

  • 储存条件

    · 12 months from date of receipt, lyophilized powder stored at -20 to -80℃.
    · 3 months, -20 to -80℃ under sterile conditions after reconstitution.
    · 1 week, 2 to 8℃ under sterile conditions after reconstitution.
    · Please avoid repeated freeze-thaw cycles.

  • 文献引用

    1.Ji Y, Gao Q, Ma Y, Wang F, Tan X, Song D, Hoo RLC, Wang Z, Ge X, Han H, Guo F, Chang J. An MMP-9 exclusive neutralizing antibody attenuates blood-brain barrier breakdown in mice with stroke and reduces stroke patient-derived MMP-9 activity. Pharmacol Res. 2023 Apr;190:106720.
    2.Gough PJ, Gomez IG, Wille PT, Raines EW. Macrophage expression of active MMP-9 induces acute plaque disruption in apoE-deficient mice. J Clin Invest. 2006 Jan;116(1):59-69.

背景介绍

  • Matrix metalloproteinase 9 (MMP-9), also known as 92 kDa type IV collagenase, 92 kDa gelatinase, gelatinase B (GELB), or CLG4B, is secreted by neutrophils, macrophages, and various transformed cells. It is the most complex member of the MMP family in terms of domain structure and activity regulation.Structurally, MMP-9 can be divided into five distinct domains: a prodomain that is cleaved upon activation, a gelatin-binding domain consisting of three contiguous fibronectin type II units, a catalytic domain with the zinc-binding site, a proline-rich linker region, and a carboxyl-terminal hemopexin-like domain. This enzyme degrades various substrates, including gelatin, collagen types IV and V, and elastin.MMP-9 is implicated in a variety of autoimmune diseases, such as systemic lupus erythematosus, rheumatoid arthritis, and multiple sclerosis, and is considered a potential therapeutic target.

操作步骤

  • 实验方法

    实验原理：检测MMP-9蛋白切割Mca-PLGL-Dpa-AR-NH2的能力。

    实验材料

    1.检测缓冲液：50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij 35, pH 7.5 (TCNB)

    2.MMP-9 His Tag Protein, Mouse

    3.p-aminophenylmercuric acetate (APMA) (Aladdin, Catalog # P649286)

    4.底物：MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (R&D, ES001)

    5.标准品：MCA-Pro-Leu-OH (Shyuanye, T77046)

    6.96 ELISA Removable Plate, Black, High binding (GENEVER, Catalog # GMO2-96H)

    7.读板器（PerkinElmer，激发波长/发射波长=320 nm/405 nm）

    实验步骤

    1.在含1mM APMA的检测缓冲液中，将MMP-9蛋白稀释至100 μg/mL，37°C孵育1小时，以激活MMP-9蛋白。

    2.用检测缓冲液将激活后的MMP-9蛋白稀释至0.8, 0.4, 0.2 μg/mL。

    3.在缓冲液中将底物稀释至20 μM。

    4.在黑色孔板中加入50 μL稀释后的MMP-9蛋白（来自步骤 2），然后加入50 μL 20 μM的底物以启动反应。需设置仅含底物和测定缓冲液的孔作为空白对照。

    5.在320 nm激发波长和405 nm发射波长下，以动力学模式（60秒/循环，20个循环）读取荧光信号。

    6.计算特异性活性：

    Specific Activity (pmol/min/µg) =

    Adjusted Vmax (RFU/min) x Conversion Factor (pmol/RFU)

    amount of enzyme (µg)


    标准曲线

    1.用检测缓冲液将标准品稀释至10 μM，并进行系列稀释。

    2.将100 μL系列稀释的标准品和空白（检测缓冲液）加入96孔板的相应孔中。标准曲线的浓度范围为每孔1000、500、250、125、62.5、31.25、15.625 pmol。

    3.分别在激发波长320 nm和发射波长405 nm下，以终点模式读取荧光信号。

    4.以MCA-Pro-Leu-OH的pmol数为纵坐标(y)，以扣除空白后的相对荧光单位 为横坐标 (x)，进行线性回归分析。

  • 电泳

    • 1μg (R: reducing condition, N: non-reducing condition).

  • 体积排阻色谱(SEC-HPLC)

    • The purity of MMP-9 His Tag Protein, Mouse is more than 90 % as determined by SEC-HPLC.

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