Furin/PCSK3 His Tag protein, Mouse

FUR,PCSK3,SPC1,PACE

Datasheet COA

价格￥4,280.00 （供应商现货： 3-5个工作日）

货号 UA011277

规格

25μg100μg500μg

数量

产品介绍

产品介绍评论(0)

产品规格

物种

Mouse
分子别名

FUR,PCSK3,SPC1,PACE
Accession

P23188
表达序列

Gln25-Glu714 with His Tag at the C-Terminus
表达宿主

HEK293
分子量

72-95kDa (Reducing)
纯度

＞85% by SDS-PAGE
活性

Measured by its ability to cleave the fluorogenic peptide substrate pERTKR-AMC. The specific activity is >310 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.
· Please do not repeated freeze-thaw cycles.
文献引用

1.Fitzgerald K. Furin Protease: From SARS CoV-2 to Anthrax, Diabetes, and Hypertension. Perm J. 2020;24:20.187.
2.Dahms SO, Hardes K, Becker GL, Steinmetzer T, Brandstetter H, Than ME. X-ray structures of human furin in complex with competitive inhibitors. ACS Chem Biol. 2014 May 16;9(5):1113-8.

背景介绍

Furin, also known as Paired Basic Amino Acid Cleaving Enzyme (PACE), is a key member of the subtilisin-like proprotein convertase family. Functioning as a calcium-dependent serine protease, its structure includes an N-terminal signal peptide, a prodomain, a catalytic domain, a P-domain responsible for maintaining enzymatic activity and calcium/pH sensitivity, a transmembrane region, and a cytoplasmic tail. Primarily localized within the Golgi apparatus/endosomal system, it activates a wide range of proprotein substrates—such as growth factors, receptors, extracellular matrix proteins, and pathogen proteins (e.g., envelope proteins of HIV and influenza virus)—by specifically cleaving after the Arg-X-(Arg/Lys)-Arg sequence motif. It plays a critical role in embryogenesis, peripheral immune tolerance, and cellular homeostasis. Dysregulation of its function is closely linked to tumor progression, neurodegenerative diseases, and various pathogen infections, making it a significant therapeutic target in clinical applications.

操作步骤

实验方法
实验原理：检测Mouse Furin Protein切割荧光肽底物Pyr-Arg-Thr-Lys-Arg-AMC的能力。
实验材料
检测缓冲液：25 mM Tris，1 mM CaCl2，pH 9.0
Furin/PCSK3 His Tag protein, Mouse (UA011277)
底物：Pyr-Arg-Thr-Lys-Arg-AMC (abs45133051)
96 ELISA Removable Plate, Black, High binding (GENEVER, Catalog # GMO2-96H)
读板器（PerkinElmer，激发波长380 nm，发射波长460 nm）
实验步骤
在检测缓冲液中将Mouse Furin Protein稀释至2 µg/mL、4 µg/mL、8 µg/mL。
在检测缓冲液中将底物稀释至100 µM。
向黑色孔板中加入2 µg/mL、4 µg/mL、8 µg/mL的Mouse Furin Protein，每孔50 µL，然后通过加入50 µL的100 µM底物来启动反应。同时需设置含50 µL的检测缓冲液和50 µL的100 µM底物的空白对照组。
在激发和发射波长分别为380 nm和460 nm的情况下，以动力学模式读取10分钟。
计算特异性活性：
*底物空白校正
**使用校准标准品7-氨基-4-甲基香豆素（7-amino-4-Methyl Coumarin）推导
转换因子（pmol/RFU）推导
缓冲液：检测缓冲液
校准标准品：7-氨基-4-甲基香豆素（Enzo，Catalog # BML-KI107-0001）。
每孔加入100 µL稀释后的校准标准品，标准曲线浓度分别为1500/750/375/188/94/4724/12/6/3/0 pmol每孔。
分别在激发波长380 nm和发射波长460 nm下以终点模式读取。
7-氨基-4-甲基香豆素（pmol（y））—RFU-空白（x）的线性回归。
备注
储存条件：复溶后，分装并保存至-80°C。禁止反复冻融。