MMP-8 His Tag Protein, Cynomolgus

MMP-8,CLG1,HNC,PMNL-CL

Datasheet COA

价格￥4,200.00 （供应商现货： 3-5个工作日）

货号 UA010308

规格

100μg500μg

数量

产品介绍

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产品规格

物种

Cynomolgus
分子别名

MMP-8,CLG1,HNC,PMNL-CL
Accession

XP_005579523.2
表达序列

Phe21-Ser467, with C-terminal 10*His
表达宿主

HEK293
分子量

65-75kDa
纯度

>95% by SDS-PAGE
内毒素含量

<0.1EU/μg
标记

Unconjugated
标签

His Tag
性状

Lyophilized Powder
缓冲体系

PBS, pH7.4
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, -20 to -70 °C as supplied; 6 months, -20 to -70 °C under sterile conditions after reconstitution; 1 week, 2 to 8 °C under sterile conditions after reconstitution; Please avoid repeated freeze-thaw cycles.
文献引用

1、Hasty K A. et al. (1990) Human neutrophil collagenase. A distinct gene product with homology to other matrix metalloproteinases. J. Biol. Chem. 265: 11421-11424.
2、Blaeser J. et al. (1991) Mercurial activation of human polymorphonuclear leucocyte procollagenase. Eur. J. Biochem. 202: 1223-1230.

背景介绍

A protease is an enzyme with a proteolytic activity that facilitates the digestion of its substrates. Membrane-type I matrix metalloproteinase (MT1-MMP), a member of the broader matrix metalloproteinases (MMP) family, is involved in the regulation of diverse cellular activities. Parasite-derived products have been shown to regulate host matrix metalloproteinases (MMPs), contributing to CE pathogenesis and progressive liver fibrosis in intermediate hosts. The current study aimed to investigate the potential role of MMP1, 7, 8, and 13 in E. granulosus induced liver fibrosis.MMP-8 may affect the metastatic behavior of breast cancer cells through protection against lymph node metastasis, underlining the importance of anti-target identification in drug development. MMP-8 in the tumor may have a protective effect against lymph node metastasis.

操作步骤

实验方法
实验原理：检测MMP-8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-8 His Tag Protein, Cynomolgus
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-8蛋白稀释至100 μg/mL，37°C孵育1小时，以激活MMP-8蛋白。
2. 用检测缓冲液将激活后的MMP-8蛋白稀释至2, 1, 0.5 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-8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，10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500、250、125、62.5、31.25、15.63、7.81、3.91、1.95 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。
酶活数据：Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001).The specific activity is >800 pmol/min/µg, as measured under the described conditions.