MMP-7/PUMP1 His Tag Protein, Human

MPSL1,Matrin,Matrix metalloproteinase-7,Pump-1 protease,Uterine metalloproteinase

Datasheet COA

价格￥1,600.00 （供应商现货： 3-5个工作日）

货号 UA016004

规格

25μg100μg500μg

数量

产品介绍

产品介绍评论(0)

产品规格

物种

Human
分子别名

MPSL1, Matrin, Matrix metalloproteinase-7, Pump-1 protease , Uterine metalloproteinase
Accession

P09237-1
表达序列

Leu18-Lys267 with His Tag at the N-Terminus
表达宿主

HEK293
分子量

25-30kDa (Reducing)
纯度

＞95% by SDS-PAGE
活性

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2. The specific activity is >3000 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.Ii M, Yamamoto H, Adachi Y, Maruyama Y, Shinomura Y. Role of matrix metalloproteinase-7 (matrilysin) in human cancer invasion, apoptosis, growth, and angiogenesis. Exp Biol Med (Maywood). 2006 Jan;231(1):20-7.
2.Adachi Y, Yamamoto H, Itoh F, Hinoda Y, Okada Y, Imai K. Contribution of matrilysin (MMP-7) to the metastatic pathway of human colorectal cancers. Gut. 1999 Aug;45(2):252-8.
3.Wang FQ, So J, Reierstad S, Fishman DA. Matrilysin (MMP-7) promotes invasion of ovarian cancer cells by activation of progelatinase. Int J Cancer. 2005 Mar 10;114(1):19-31.

背景介绍

MMP-7 (Matrix Metalloproteinase-7), also known as Matrilysin or PUMP-1, is the smallest secreted zinc-dependent endopeptidase in the matrix metalloproteinase family, with a molecular weight of approximately 28 kDa and specific expression in epithelial cells. It has an extremely streamlined structure comprising only a signal peptide, pro-domain, and catalytic domain, lacking the hinge region and hemopexin-like domain commonly found in other MMPs. The HEXXHXXGXXH motif in its catalytic domain exerts proteolytic activity by chelating zinc ions, enabling it to degrade basement membrane components such as laminin, fibronectin, and type IV collagen, as well as activate inflammatory factors like pro-TNF-α and cleave E-cadherin to promote epithelial-mesenchymal transition. Under physiological conditions, it participates in tissue remodeling, wound healing, and gland development, whereas under pathological conditions, MMP-7 is highly expressed in various malignancies including gastric, colorectal, pancreatic, and renal cell carcinomas, closely correlating with poor prognosis through promoting tumor invasion, angiogenesis, and lymph node metastasis. Plasma MMP-7 level detection has demonstrated favorable diagnostic value (AUC 0.80-0.91) and independent prognostic predictive capability, and can assess pathological response to neoadjuvant therapy in pancreatic cancer patients, making it a highly promising tumor biomarker and therapeutic target.

操作步骤

实验方法
实验原理：检测MMP-7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-7/PUMP1 His Tag Protein, Human (active enzyme)
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-7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-7蛋白稀释至0.4, 0.2, 0.1 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-7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。
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。