辣根过氧化物酶（Horseradish Peroxidase, HRP）

产品包装规格：

种属：             Horseradish

表达系统：     Prokaryotic expression system、Eukaryotic expression system

标签：              N-His

同用名：          Peroxidase (POD) from Horseradish

分子量：          44 KDa

纯度：           lyophilized powder，≥340U/mg

储存条件：      1mg/ml solution (0.1M phosphate buffer, pH6.0)

备注：          Please use this product immediately after receiving it or store it in accordance with the conditions recommended in the instructions.With the extension of preservation time, the enzyme activity will decrease to a certain extent.This product is freeze-dried powder. After 2 weeks of storage at 37ºC, the enzyme activity can be maintained at least 80%.This product is only used for scientific research by professionals, shall not be used for clinical diagnosis or treatment, shall not be used for food or medicine, and shall not be stored in ordinary homes.   For your safety and health, please wear a lab coat and disposable gloves.           

储存时间：    Store at 4ºC or -20ºC away from light for at least two years. 

运输：             The product is shipped at ambient temperature.

背景：             Horseradish Peroxidase (HRP)，也称Peroxidase (POD) from Horseradish，即辣根过氧化物酶，EC号1.11.1.7 ，CAS号9003-99- 0 ，是从植物辣根(horseradish)中提取得到的一种含有血红素的氧化还原酶， 以过氧化氢或其它有机过氧化物为电子受体催 化氧化多种有机物和无机物底物。本产品的酶活性≥340U/mg。

> 本产品用途广泛。由于HRP分子量较小且稳定，并且HRP本身的糖基化修饰使其非特异性结合非常低，所以常用于标记二 抗、 一抗或Streptavidin等而用于Western blot 、ELISA、免疫组化等检测， HRP也常用于一些生化检测。

> 本产品为棕褐色粉末状的高纯度HRP。本产品呈现棕褐色是由于HRP中含有血红素辅基导致的， 并非由于纯度不高导致 的。本产品RZ≥3 ，酶活性≥340U/mg，溶解度≥10mg/ml 。RZ (Reinheitszahl)定义： HRP分子中的血红素辅基在403nm波长处 有最大吸收峰，而去辅基的HRP在275nm波长处有最大吸收峰， A403/A275比值即为RZ值， RZ值仅反映了血红素含量， 并 不代表酶活性的高低， 高纯度HRP的RZ值应在3.0左右。酶活力单位定义： 20ºC ，pH6.0条件下反应20秒，氧化邻苯三酚 (pyrogallol)得到1.0mg红棓酚(purpurogallin)所需的酶量定义为一个活力单位。

> HRP是一种含四个二硫键的单链多肽， 由无色的酶蛋白和棕色的铁卟啉(ferroprotoporphyrin)结合而成的糖蛋白，分子量约 44kDa，包括多肽链(33890Da) 、血红素(约700Da)和糖类物质(约9400Da)，其中碳水化合物含量达18% 。不同HRP同工酶中 包含的糖类物质有所不同，主要包括半乳糖、阿拉伯糖、木糖、海藻糖、甘露糖、甘露糖胺和半乳糖胺等。目前已发现至 少有7种HRP同工酶，不同同工酶的等电点大约在3.0-9.0之间。

> HRP的作用机理：HRP与过氧化氢结合，得到的[HRP-H2O2]复合物能够使多种氢供体底物发生氧化，而供氢体底物大多为 无色的还原型显色底物(chromogenic  substrate，如DAB 、TMB 、ABTS等)、化学发光底物(chemiluminescent  substrate，如 luminol等)和荧光底物(fluorogenic  substrate，如Amplex Red等) ，通过反应可生成氧化型的有色染料、化学发光或荧光产 物，最终通过适当仪器进行定性或定量检测。

> HRP常通过二硫键介导的戊二醛交联、过碘酸氧化法及氨基和巯基直接介导的交联等多种方式对抗体进行标记，HRP与β- 半乳糖苷酶(β-galactosidase)和碱性磷酸酶(alkaline phosphatase)相比具有更小的分子量和更高的稳定性， 因此HRP是三者中 最为理想的抗体标记物。

> HRP最适pH在6.0-6.5之间， 在pH5.0-9.0之间都处于稳定状态。 HRP常见的抑制剂有叠氮化钠、氰化物、 L-胱氨酸、重铬酸 盐、羟胺类物质、钒酸酯、 P-氨基苯甲酸和Cd2+ 、Co2+ 、Cu2+ 、Fe2+ 、Mn2+ 、Ni2+ 、Pb2+等一些金属离子。

Horseradish Peroxidase (HRP), also known as Peroxidase (POD) from Horseradish, EC No. 1.11.1.7, CAS No. 9003-99-0, A heme-containing oxidoreductase extracted from the plant horseradish, which uses hydrogen peroxide or other organic peroxides as electron acceptors to catalyze the oxidation of various organic and inorganic substrates. The enzyme activity of this product is ≥340U/mg.

> This product is versatile. Due to the small and stable molecular weight of HRP, and the glycosylation modification of HRP itself, its non-specific binding is very low, so it is often used to label secondary antibody, primary antibody or Streptavidin, and for Western blot, ELISA, immunohistochemical and other detection. HRP is also often used for some biochemical detection.

> This product is high purity HRP in tan powder form. The brown color of this product is due to the heme cogroup contained in HRP, not due to low purity. RZ≥3, enzyme activity ≥340U/mg, solubility ≥10mg/ml. RZ (Reinheitszahl) definition: The maximum absorption peak of the heme cogroup in HRP was found at the wavelength of 403nm, while the maximum absorption peak of the uncogroup HRP was found at the wavelength of 275nm. The ratio of A403/A275 was the RZ value, and the RZ value only reflected the heme content, but did not represent the level of enzyme activity. The RZ value of high-purity HRP should be around 3.0. Enzyme activity unit definition: 20ºC, pH6.0 reaction for 20 seconds, pyrogallol oxidation pyrogallol to obtain 1.0mg purpurogallin was defined as a unit of activity.

> HRP is a single chain polypeptide containing four disulfide bonds. It is a glycoprotein formed by the combination of colorless enzyme protein and brown ferroprotoporphyrin, with a molecular weight of about 44kDa, including polypeptide chain (33890Da), heme (about 700Da) and carbohydrate substance (about 9400Da). The carbohydrate content is 18%. The sugars contained in different HRP isoenzymes were different, mainly including galactose, arabinose, xylose, trehalose, mannose, mannose amine and galactosamine. At least 7 HRP isoenzymes have been found, and the isoelectric points of different isoenzymes are about 3.0-9.0.

> HRP mechanism: HRP combined with hydrogen peroxide, the obtained [HRP-H2O2] complex can oxidize a variety of hydrogen donor substrates, and most of the hydrogen donor substrates are colorless chromogenic substrates. Such as DAB, TMB, ABTS, etc.), chemiluminescent substrate (such as luminol, etc.) and fluorogenic substrate (such as Amplex Red, etc.), Through the reaction, oxidized colored dyes, chemiluminescence or fluorescence products can be generated, which are finally tested qualitatively or quantitatively by appropriate instruments.

> HRP usually labels antibodies through a variety of ways, such as glutaraldehyde crosslinking mediated by disulfide bond, periodate oxidation and crosslinking directly mediated by amino and sulfhydryl groups. HRP has a smaller molecular weight and higher stability than β-galactosidase and alkaline phosphatase, making it the most ideal antibody marker of the three.

> The optimum pH of HRP is between 6.0-6.5, and the pH is stable between 5.0-9.0. The common inhibitors of HRP include sodium azide, cyanide, L-cysteine, bichromate, hydroxylamines, vanadates, P-aminobenzoic acid and some metal ions such as Cd2+, Co2+, Cu2+, Fe2+, Mn2+, Ni2+, Pb2+.
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