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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, Human

Type V Collagenase,Gelatinase B,EC 3.4.24.35,MANDP2,Matrix Metalloproteinase 9,CLG4B,MMP9,Matrix Metallopeptidase 9,Matrix Metalloproteinase 9 (Gelatinase B,92kDa Gelatinase,92kDa Type IV Collagenase),GELB,Matrix Metalloproteinase-9,92 KDa Type IV Collagenase,Macrophage Gelatinase,92 kDa gelatinase,MMP-9,Matrix Metallopeptidase 9 (Gelatinase B,EC:3.4.24.35,EC 3.4.24

价格 ￥1,600.00 （ 供应商现货 ： 3-5个工作日）
货号 UA011205
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

    Human

  • 分子别名

    Type V Collagenase，Gelatinase B，EC 3.4.24.35，MANDP2，Matrix Metalloproteinase 9，CLG4B，MMP9，Matrix Metallopeptidase 9，Matrix Metalloproteinase 9 (Gelatinase B， 92kDa Gelatinase， 92kDa Type IV Collagenase)，GELB，Matrix Metalloproteinase-9，92 KDa Type IV Collagenase，Macrophage Gelatinase，92 kDa gelatinase，MMP-9，Matrix Metallopeptidase 9 (Gelatinase B， 92kDa Gelatinase， 92kDa Type IV Collagenase)，EC:3.4.24.35，EC 3.4.24

  • Accession

    P14780-1

  • 表达序列

    Ala20-Pro469 with His Tag at C-Terminus

  • 表达宿主

    HEK293

  • 分子量

    55-72kDa (Reducing)

  • 纯度

    ＞95% by SDS-PAGE & SEC-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

  • 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.St-Pierre Y, et al. (2003) Emerging features in the regulation of MMP-9 gene expression for the development of novel molecular targets and therapeutic strategies. Curr Drug Targets Inflamm Allergy. 2(3): 206-15.
    2.St-Pierre Y, et al. (2004) Regulation of MMP-9 gene expression for the development of novel molecular targets against cancer and inflammatory diseases. Expert Opin Ther Targets. 8(5): 473-89.
    3.Chakrabarti S, et al. (2005) Matrix metalloproteinase-2 (MMP-2) and MMP-9 in pulmonary pathology. Exp Lung Res. 31(6): 599-621.

背景介绍

  • Matrix metalloproteinases (MMPs) are neutral proteases involved in extracellular matrix (ECM) breakdown and remodeling during physiological processes like morphogenesis, angiogenesis, and tissue repair, as well as in pathological conditions such as inflammation, arthritis, and cancer. MMP9, or 92-kDa gelatinase B/type IV collagenase, is secreted by neutrophils, macrophages, and transformed cells. It is critical for tissue remodeling in inflammatory processes and is a key product of macrophages and neutrophils. MMP9 is also released by lymphocytes and stromal cells in response to inflammatory cytokines or integrin-mediated signals. Excessive MMP9 production is linked to tissue damage and degenerative disorders, making its regulation a focus for therapeutic development. Its overexpression in cancer and inflammatory conditions highlights the importance of understanding the mechanisms controlling its expression to identify potential therapeutic targets.

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, Human

    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4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).

  • 体积排阻色谱(SEC-HPLC)

    • The purity of MMP-9 His Tag Protein, Human is greater than 95% as determined by SEC-HPLC.

  • ELISA

    • Immobilized MMP-9 His Tag Protein, Human (Cat. No. UA011205) at 2.0μg/mL (100μL/well) can bind MMP9 Recombinant Rabbit mAb (SDT-368-16) (Cat. No. S0B2297) with EC50 of 0.95-1.48ng/mL.

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