FAP His Tag Protein, Human (active dimer)

FAPA,SIMP,DPPIV,FAP alpha

价格￥3,000.00 （ 1-2周）

货号 UA010165

规格

25μg100μg500μg

数量

产品介绍

产品介绍引用文献(1) 评论(0)

产品规格

物种

Human
分子别名

FAPA,SIMP,DPPIV,FAP alpha
Accession

Q12884
表达序列

Leu26-Asp760, with N-8*His
表达宿主

HEK293
分子量

80-90kDa
纯度

>95% by SDS-PAGE
内毒素含量

<0.1EU/μg
活性

Measured by its ability to convert the substrate benzyloxycarbonyl-Gly-Pro-7-amido-4-methylcoumarin (Z-GP-AMC) to Z-Gly-Pro and 7-amino-4-methylcoumarin (AMC). The specific activity is >10000 pmol/min/μg, as measured under the described conditions.
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, 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、Cheng J D. et al. (2005) Fibroblast activation protein overexpression confers an enhanced adherence and motility phenotype. Cancer Res. 65(1): 1326.
2、Kraman M. et al. (2010) Suppression of Antitumor Immunity by Stromal Cells Expressing Fibroblast Activation Protein-α. Science. 330(6005): 827-830.
3、Bauer S. et al. (2006) Fibroblast activation protein is expressed by rheumatoid myofibroblast-like synoviocytes. Arthritis Res Ther. 8(6): R171.

背景介绍

Neuropeptide-associated fibroblast activation protein (FAP) is a homodimeric integral membrane gelatinase belonging to the serine protease family. It is selectively expressed in reactive stromal fibroblasts of epithelial cancers, granulation tissue of healing wounds, and malignant cells of bone and soft tissue sarcomas. This protein is thought to be involved in the control of fibroblast growth or epithelial-mesenchymal interactions during development, tissue repair, and epithelial carcinogenesis. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. Neuropeptide-associated fibroblast activation protein (FAP) may be an important risk factor for neurovascular metastasis in hepatocellular carcinoma. Analysis of The Cancer Genome Atlas (TCGA) database showed that FAP mRNA was highly expressed in most human tumor tissues. The HPA database then verified that FAP was highly expressed in tumor tissues following protein translation. Survival analysis then showed that the level of FAP expression significantly affected the overall survival (OS), progress free interval (PFI), and disease specific survival (DSS) of patients with hepatocellular carcinoma.

操作步骤

实验方法
实验原理：检测FAP切割荧光肽底物Z-GP-AMC的能力。
实验材料
1. 检测缓冲液：50 mM Tris, 1 M NaCl, 1 mg/mL BSA, pH 7.5
2. FAP His Tag Protein, Human
3. 底物：Z-Gly-Pro-AMC (Bachem, Catalog # I-1145)
4. 96 ELISA Removable Plate, Black, High binding (GENEVER, Catalog # GMO2-96H)
5. 读板器（PerkinElmer，激发波长380 nm，发射波长460 nm）
实验步骤
1. 用检测缓冲液将FAP蛋白稀释至浓度0.4 μg/mL、0.2 μg/mL。
2. 用检测缓冲液将底物稀释至浓度100μM。
3. 向酶标板中分别加入50 μL浓度为0.4 μg/mL、0.2 μg/mL的FAP稀释液，再加入50 μL 100 μM底物溶液启动反应；同时设置底物空白对照组：加入50 μL检测缓冲液+50 μL底物。
4. 采用顶部读板模式，在激发波长380 nm、发射波长460 nm条件下，动力学模式连续读取荧光信号5分钟。
5. 计算特异性酶活性：
Specific Activity (pmol/min/µg) =
Adjusted Vmax* (RFU/min) x Conversion Factor (pmol/RFU)**
amount of enzyme (µg)
*底物空白校正
**本数值由校准标准品7-氨基-4-甲基香豆素（7-AMC，品牌Enzo，货号BML-KI107-0001）计算得出

电泳
- 1μg (R: reducing conditions, N: non-reducing conditions).
体积排阻色谱(SEC-HPLC)
- The purity of Human FAP, His Tag (Cat. No. UA010165) is more than 95% and the molecular weight of this protein is around 219 kDa verified by SEC-MALS.
ELISA
- Immobilized FAP His Tag, Human (Cat. No. UA010165) at 1.0μg/mL (100μL/well) can bind FAP Recombinant Rabbit mAb (SDT-R132) (Cat. No. S0B2166) with EC50 of 2.15-3.34ng/mL.
- Immobilized FAP His Tag Protein, Human (Cat. No. UA010165) at 1.0μg/mL (100μL/well) can bind Sibrotuzumab with EC50 of 0.37-0.67ng/mL.

引用文献(1)