ACPP/ACP3 His Tag Protein, Human

ACP3,PAP,5'-NT,ACP-3,Prostatic Acid Phosphatase

价格￥1,200.00 （供应商现货： 3-5个工作日）

货号 UA040045

规格

10μg50μg100μg500μg

数量

产品介绍

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产品规格

物种

Human
分子别名

ACP3,PAP,5'-NT,ACP-3,Prostatic Acid Phosphatase
Accession

P15309
表达序列

Lys33-Asp386, with C-terminal His Tag
表达宿主

HEK293
分子量

45-50kDa (Reducing)
纯度

＞95% by SDS-PAGE
内毒素含量

<0.1EU/μg
活性

Measured by its ability to cleave a substrate, p-Nitrophenyl phosphate (pNPP). The specific activity is >300,000 pmol/min/μg.
标记

Unconjugated
标签

His Tag
性状

Lyophilized Powder
缓冲体系

PBS, 5% Trehalose, 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.

背景介绍

Prostatic acid phosphatase (PAP, or ACPP), also known as prostatic specific acid phosphatase (PSAP), is an enzyme produced by the prostate. As a non-specific phosphomonoesterase, Prostatic acid phosphatase is synthesized and secreted into seminal plasma under androgenic control. The enzyme is a dimer of molecular weight around 100 kDa. Prostatic Acid Phosphatase (ACPP) catalyzes the hydrolysis of a variety of phosphate monoesters, including phosphorylated proteins. The activity optimum of ACPP is in the pH range of 4-6, and the activity is inhibited by L(+)-tartrate. ACPP expression levels are highest in the prostate, with much lower expression in most other tissues. ACPP is a type I integral membrane protein of the plasma membrane and lysosomes, and a secreted form also exists. The concentration of ACPP is elevated in the circulation of prostate cancer patients, making the enzyme a marker for the progression of prostate cancer. Cellular ACPP has been shown to be a protein tyrosine phosphatase. Protein substrates include the epidermal growth factor receptor and HER-2. Cellular ACPP is considered to function as a tumor suppressor.

操作步骤

实验方法
实验原理：检测ACPP His Tag Protein, Human裂解底物pNPP的能力。
实验材料
1. 检测缓冲液：50 mM NaOAc, pH 4.5
2. ACPP His Tag Protein, Human
3. 底物：p-Nitrophenyl phosphate (Sigma-Aldrich, Catalog # 241326)
4. NaOH, 0.2 M in deionized water
5. 标准品：4-Nitrophenol (Sigma-Aldrich, Catalog #N2765)
6. 96 Well Clear Plate (BIOFIL, Catalog#011096)
7. 读板器（PerkinElmer）
实验步骤
1. 在检测缓冲液中将ACCP蛋白稀释至0.4 µg/mL。
2. 在检测缓冲液中将底物稀释至2 mM。
3. 在96孔板中加入50 µL稀释后的ACCP蛋白（步骤1）。然后加入50 µL的2 mM底物以启动反应。同时设置仅含检测缓冲液和底物的对照组。
4. 室温下孵育5分钟。
5. 加入 100 μL 0.2 M NaOH 以终止反应并显色。
6. 在终点模式下于 410 nm（顶读）处读取吸光度。
7. 计算特异性活性：
Specific Activity (pmol/min/µg) =
Adjusted Vmax (OD)x Conversion Factor (pmol/OD)
Incubation time(min) x amount of enzyme (μg)