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P8375

Sigma-Aldrich

过氧化物酶 来源于辣根

Type VI, essentially salt-free, lyophilized powder, ≥250 units/mg solid (using pyrogallol)

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别名:
HRP, 检测酶, 过氧化物酶, 供体：过氧化氢氧化还原酶, 辣根过氧化物酶
CAS号:
9003-99-0
EC 号:
1.11.1.7 (BRENDA, IUBMB)
EC 号:
232-668-6
MDL编号:
MFCD00071339
eCl@ss:
32160410
NACRES:
NA.54

类型

Type VI

质量水平

300

形式

essentially salt-free, lyophilized powder

specific activity

≥250 units/mg solid (using pyrogallol)

分子量

~44 kDa

溶解性

0.1 M phosphate buffer: soluble 10 mg/mL, clear, orange to red (pH 6.0)

吸光度比值

RZ 2.5-4.0

application(s)

diagnostic assay manufacturing

储存温度

2-8°C

InChI

1S/H2O3/c1-3-2/h1-2H

InChI key

JSPLKZUTYZBBKA-UHFFFAOYSA-N

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相关类别

一般描述

辣根过氧化物酶是从辣根（Amoracia rusticana）中分离出来的，属于过氧化物酶的铁原卟啉基团。HRP是含有四个二硫键的单链多肽。它是一种含有18%碳水化合物的糖蛋白。碳水化合物由半乳糖、阿拉伯糖、木糖、岩藻糖、甘露糖、甘露糖胺和半乳糖胺组成，具体取决于特定同工酶。其分子量（约44 kDa）包括多肽链（33,890道尔顿）、氯化血红素加Ca2+ （约700道尔顿）和碳水化合物（约9,400道尔顿）。至少存在7种HRP同工酶。 辣根过氧化物酶同工酶的等电点范围为3.0-9.0。

应用

该酶已用于开发基于热稳定性大豆过氧化物酶的生物传感器，通过氧化还原传导水凝胶将酶交联并“电连接”到玻璃碳电极上。它还用于测定低密度脂蛋白的氧化。
辣根过氧化物酶（HRP）分离自辣根（紫穗槐），属于过氧化物酶的铁原卟啉基团。 它被用作western blot、ELISA和免疫组化等生化应用。 辣根过氧化物酶用于扩增弱信号并提高靶标分子如蛋白的可检测性。 产品P8375，VI型，是一种基本不含盐的冻干粉。 它常用于测定溶液中葡萄糖和过氧化物的量。 它已用于研究颈脊髓神经元的感觉输入。

生化/生理作用

HRP易与过氧化氢（H2O2）结合，而所得的[HRP-H2O2]络合物可氧化各种氢供体。最适pH为6.0-6.5而酶在5.0-9.0的pH范围内最为稳定。HRP可以通过几种不同的方法与抗体结合，包括戊二醛，高碘酸盐氧化，通过二硫键，以及通过氨基和巯基定向交联剂。它比酶标记β-半乳糖苷酶和碱性磷酸酶更小且更稳定，因此是最理想的标记。此外，它的糖基化会导致更低低的非特异性结合。
当与底物一起孵育时，辣根过氧化物酶可产生标记分子的有色、荧光或发光衍生物，从而可实现定量。 辣根过氧化物酶已被证明可略微降低cydAB突变体中的抑制水平。 已知的抑制剂包括叠氮化钠、氰化物、L-胱氨酸、重铬酸盐、亚乙基硫脲、羟胺、硫化物、钒酸盐、对氨基苯甲酸以及Cd2+、Co2+、Cu2+、Fe3+、Mn2+、Ni2+以及Pb2+离子。

单位定义

一个连苯三酚单元将在20℃、pH 6.0条件下，在20秒内从连苯三酚形成1.0mg的红棓酚。

分析说明

RZ(Reinheitszahl)是指溶于去离子水中的酶(0.5–1.0 mg/ml)的吸光度比值A403/A275 。 它是血红素含量的量度，而不是酶活性。 即使是RZ值较高的制剂也可能具有较低的酶活性。

其他说明

有关过氧化物酶的更多信息，请访问 www.sigma-aldrich.com/enzymeexplorer。

象形图

Health hazard
GHS08

警示用语：

Danger

危险声明

H317 - H334

危险分类

Resp. Sens. 1 - Skin Sens. 1

储存分类代码

11 - Combustible Solids

WGK

WGK 3

个人防护装备

Eyeshields, Gloves, type N95 (US)

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索取COA

  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  3. How do I convert units of Peroxidase from horseradish, Product P8375, to solid?

    The activity listed on the Certificate of Analysis for each lot can be used to convert from units to milligrams of solid.

  4. What is the optimal pH range of Peroxidase from horseradish, Product P8375?

    The optimal pH range for product P8375 is 6.0 - 6.5.

  5. What can you tell me about the stability of Peroxidase from horseradish, Product P8375, (as is, in suspension and in solution)?

    This product is stable as a powder, kept refrigerated. See the certificate of analysis for a lot-specific recommended retest date. It is also stable as a crystalline suspension of 3.2 M (NH4)2SO4 solution containing potassium phosphate buffer, pH 6.0 (see Product No. P6140). It is reasonably stable in solution in phosphate buffer. We have found that at 10 mg/mL in 0.1 phosphate buffer, pH 6.0, solutions kept frozen at -20°C lose less than 2% of their activity per week.

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    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

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实验方案

Spectrophotometric Determination of Reinheitszahl (RZ) for Peroxidase (EC 1.11.1.7)

Reinheitszahl (RZ) is the ratio of absorbance due to hemin (A403, Soret region) to absorbance due to protein (A275).

Enzymatic Assay of Peroxidase (EC 1.11.1.7) 2,2'-Azino-bis(3-Ethylbenzthiazoline-6-Sulfonic Acid) as a Substrate

To standardize a procedure for the assay of Peroxidase using 2,2'-Azino-bis(3-Ethylbenzthiazoline-6-Sulfonic Acid) as a substrate.

Enzymatic Assay of Peroxidase (EC 1.11.1.7)

This procedure is for the determination of Peroxidase enzymatic activity using Pyrogallol as the substrate.

过氧化物酶的酶学测定（EC 1.11.1.7）

该实验程序用于使用连苯三酚作为底物测定过氧化物酶的酶活性。

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