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品种反应性 主要应用 检测方法 H ACT Colorimetric 描述 产品目录编号 ECM600 品牌系列 Chemicon®
商名 - Chemicon
描述 uPA Activity Assay Kit 概述 Urokinase-type Plasminogen Activator (uPA) is a 52 kDa serine protease which has been implicated in a number of physiological and pathological processes, including tissue remodeling1, angiogenesis2, fibrinolysis and tumor spread. When bound to its cell surface receptor, uPA is converted from the single chain pro-form uPA to the active 2-chain HMW-uPA. uPA has been shown to play a role in basement membrane degradation, via a cascade involving activation of plasminogen and the matrix metalloproteinases3. Inhibitors of uPA have been shown to slow primary tumor growth and metastasis4-7.
The CHEMICON uPA Activity Assay Kit provides a quick, efficient and sensitive system for evaluation of uPA activity and for screening of uPA inhibitors. The assay is colorimetric and does not require radioactivity or fluorescence equipment. The assay is sensitive over a range of 0.05-50 units of uPA activity.
Test Principle:
The CHEMICON uPA Activity Assay Kit utilizes a chromogenic substrate, which is cleaved by active uPA. Addition of this substrate to a uPA-containing sample results in a colored product, detectable by its Optical Density at 405nm (OD405).
Application:
The CHEMICON uPA Assay Kit is ideal for measurement of uPA activity in purified preparations and cell culture, as well as in serum where pathological conditions such as sepsis exist. The assay is also useful for screening inhibitors of the enzymatic activity of uPA.
Each CHEMICON uPA Activity Assay Kit contains sufficient reagents for the evaluation of 96 samples, including uPA from human urine as a positive control. Duplicate or triplicate samples are suggested.
The CHEMICON uPA Activity Assay Kit is intended for research use only; not for diagnostic or therapeutic applications.产品信息 部件 - Assay Buffer, 10X: (Part No. 90091) One 5 mL bottle.
- uPA Positive Control: (Part No. 90058) One lyophilized vial, 1000 units, of uPA from human urine.
- Chromogenic Substrate: (Part No. 90057) One 5 mg bottle of Tripeptide with pNA group.
检测方法 Colorimetric 应用 应用 The uPA Activity Assay Kit provides a quick, efficient & sensitive system for evaluation of uPA activity & for screening of uPA inhibitors. 主要应用 - Activity Assay
应用说明 Notes on uPA activity assay sample preparation.
Plasma: Perform venepuncture of the cubial vein, with minimal stasis after a rest period of at least 10min. Discard the first few milliliters of blood. Collect nine volumes of blood in one volume of cold sodium citrate, 0.11 mol/L, in a polycarbonate tube, mix gently and place on ice. Subsequently, prepare platelet-poor plasma by centrifugation in a refrigerated centrifuge (30 min, 2000g). Snap-freeze plasma samples of 0.3 mL to 0.6 mL and store at -60°C. Before use, thaw plasma rapidly in a water bath at 37°C and put on ice.
Tissue: There are numerous publications for the extraction of uPA from tissues. A suitable buffer for extraction of uPA from the membrane fraction requires Triton X100. A non-detergent extract (also called cytosolic fraction) does not require Triton, however the membrane fraction is removed by ultracentrifugation, so uPA activity associated with the membrane fraction is bypassed by measuring only the cytosols. Below are some references that describe extraction procedures for uPA.
Cancer Res. (1987). 47(17):4654-4657.
J. Bone Miner Res. (1991). 6(10):1081-1090.
Cancer Res. (1994). 54(10):2527-2530.
Br. J. Cancer (1996). 74(8):1168-1174.
Cell culture: Isolate cell culture supernatant and use directly.生物信息 品种反应性 Human Entrez基因编号 - NM_002658.2
Entrez基因汇总 This gene encodes a serine protease involved in degradation of the extracellular matrix and possibly tumor cell migration and proliferation. A specific polymorphism in this gene may be associated with late-onset Alzheimer disease and also with decreased affinity for fibrin-binding. The protein encoded by this gene converts plasminogen to plasmin by specific cleavage of an Arg-Val bond in plasminogen. This gene's proprotein is cleaved at a Lys-Ile bond by plasmin to form a two-chain derivative in which a single disulfide bond connects the amino-terminal A-chain to the catalytically active, carboxy-terminal B-chain. This two-chain derivative is also called HMW-uPA (high molecular weight uPA). HMW-uPA can be further processed into LMW-uPA (low molecular weight uPA) by cleavage of chain A into a short chain A (A1) and an amino-terminal fragment. LMW-uPA is proteolytically active but does not bind to the uPA receptor. 基因符号 - Urokinase-type plasminogen activator chain B].
- PLAU
- UPA
- URK
- u-PA
- uPA
- ATF
- EC 3.4.21.73 [Contains: Urokinase-type plasminogen activator long chain A
- Urokinase-type plasminogen activator short chain A
UniProt编号 - P00749
UniProt汇总 FUNCTION: SwissProt: P00749 # Specifically cleave the zymogen plasminogen to form the active enzyme plasmin.
SIZE: 431 amino acids; 48525 Da
SUBUNIT: Found in high and low molecular mass forms. Each consists of two chains, A and B. The high molecular mass form contains a long chain A which is cleaved to yield a short chain A. Binds LRP1B; binding is followed by internalization and degradation. Interacts with MRC2. Interacts with PLAUR.
SUBCELLULAR LOCATION: Secreted.
TISSUE SPECIFICITY: Expressed in the prostate gland and prostate cancers.
PTM: Phosphorylation of Ser-158 and Ser-323 abolishes proadhesive ability but does not interfere with receptor binding.
SIMILARITY: SwissProt: P00749 ## Belongs to the peptidase S1 family. & Contains 1 EGF-like domain. & Contains 1 kringle domain. & Contains 1 peptidase S1 domain.理化信息 灵敏度 - Positive Control: Specific activity: =100,000 units/mg protein. One unit is defined as the amount of enzyme equal to an international standard tested by the fibrinolytic method of Johnson, et al. {Johnson, A.J., et al. 1969. Thromb. Diath. Haemorrh. 21, 259.}
~10 μg of standard is supplied in ECM600
产品使用声明 使用声明 - Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
储存和货运信息 存储条件 Store kit materials at -20°C for up to 6 months. After reconstitution, store uPA positive control at -70°C and substrate at 2 - 8°C. 包装信息 数量 1 kit
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