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| 别名 | Donor: hydrogen-peroxide oxidoreductase；Horseradish peroxidase；HRP POD |
| 英文名称 | Peroxidase(POD) |
| CAS | No.9003-99-0 |
| 储存条件 | -20℃，有效期5年 |
| 外观(性状) |
褐色结晶性冻干粉状
|
| 来源 | From horseradish |
| 酶活/效价 | 233-300units/mg solid （以实际收到的标签酶活为准） |
| 溶解性 | 10 mg/mL in water |
| 规格 | 10mg 100mg 1g |
| 单位 | 支 |
说明：POD是一种具有氧化还原酶性质的血红素蛋白。用于生化研究，是酶标记法所用的一种主要酶。也用于生物液体中 葡萄糖和半乳糖的测定
HRP是一种分子量达44 000的糖蛋白，由无色的酶蛋白和深棕色的铁卟啉结合而成，中性糖和氨基糖约占18％，主要有甘露糖、木糖、阿拉伯糖和己糖胺等。每一个HRP分子中含一个氯化血红素IX作辅基，该辅基在403nm波长处有最大吸收峰，而去辅基的酶蛋白在275nm波长处有最大吸收。HRP在403nm的OD值与275nm的OD值之比，也就是所谓的RZ(Reinheits Zahl)值。RZ值仅说明血红素基团在HRP中的含量，并非表示HRP制剂的真正纯度，而且RZ值高的HRP制剂，并不意味着酶活性也高。但可以一纯酶溶液在10mm光径403nm波长下的吸光度来计算酶浓度[1％(W/V)酶溶液吸光度为22.5，浓度为227umol/L]。纯HRP干燥贮存于–20℃可保持稳定，使用1.36 mol/L甘油、10mmol/L磷酸钠、30umol/L牛血清白蛋白和20umol/L细胞色素C(pH 7.4)溶液作为基质冷冻保存，可使酶结合物稳定数年。HRP对热及有机溶剂的作用比较稳定，用甲苯与石蜡切片处理或用纯乙醇或l0％甲醛水溶液固定做冷冻切片，均不能使其活性改变。氰化物或硫化物在10–5～10–6mol/L浓度时具有可逆性地抑制HRP的作用；氟化物、叠氮化合物或羟胺仅在高于10-3mol／L浓度时抑制HRP；HRP还可被羟甲基过氧化氢不可逆地抑制。强酸也是HRP的强烈的抑制剂。
因此，在酶免疫测定常选用上述的某些化合物如氟化钠、叠氮钠和强酸等作为酶反应的终止剂。此外，在配制酶免疫测定的稀释缓冲液时，为防止酶失活，应避免使用叠氮钠作防腐剂。HRP的同工酶主要可分为三种类型：①含糖量高的酸性同工酶。②等电点接近于中性(或微碱)的含糖量相对较低的同工酶。③含糖量低的碱性(PI>11)同工酶。酶免疫试验中所使用的HRP，以PI为8.7～9.0的所谓“C”同工酶为主要组成成分，其他同工酶的活性则很低。“C”同工酶的共价结构由2个密切接近的区域组成，血红素基团则位于其间而成夹心结构，糖链以8个不同的部位结合于多肽。天然的酶所带的纯电荷极少；无游离的。—氨基，仅有2个可测出的组氨酸，6个赖氨酸似乎全被糖链外壳所遮盖。因此，HRP一般仅有1～2个可用于耦联的氨基。
根据HRP的催化特性，ELISA中一般使用过氧化氢(H2O2)作为HRP底物之一，在供氢体(即色原底物)存在时，HRP与H2O2的反应迅速而又专一。由图4可见，HRP被H2O2二价地氧化形成复合物I，而复合物I又可与氢供体的二步连续的单价相互作用而还原至起始状态。复合物Ⅱ是被氧化的带一个电子的中间产物，当H2O2过量，由于复合物Ⅲ或Ⅳ的形成，酶活性受抑制(以后补上)。30％的H2O2并不稳定。由于H2O2既为HRP的底物，也为其抑制剂，因而ELISA要想得到满意的测定结果，H2O2必须限定在一定的浓度范围内，终浓度通常为2～6 mmol/L。然而在实际研究工作中，一般很少注意这一点。大多数研究者所使用的H2O2的浓度常较理想反应所需的量大2～4倍。吸附于固相的HRP较游离的HRP更易受过量H2O2的抑制。如果30％H2O2贮存液浓度经测定证实确为30％，则稀释10 000—12 000倍常是较为理想的底物。H2O2的摩尔消光系数为10mm光径240nm波长下为43.6，因此，可通过这种方式来检测H2O2工作溶液的浓度。
固相ELISA中，当温度高于-20℃时，HRP活性常较低，在底物溶液中加入非离子去垢剂聚山梨醇-20或TritonX-100可延迟HRP的失活，且可使反应温度加大，但非离子去垢剂的这种酶活性保护效应依氢供体的不同而有差异。如以2,2’-联氮-双-[3-乙基苯并噻唑啉]-6-磺酸(ABTS)为氢供体，则只能保护20％的酶活性，而以邻联茴香胺(ODA)为氢供体时，酶活性的保护提高至90％。
HRP之所以是迄今为止在ELISA中应用最为广泛的标记用酶，主要是因为其一方面易于提取，价格相对低廉；另一方面性质稳定，耐热及有机溶剂的作用，与抗原或抗体偶联后，活性很少受损失。
备注：产品信息可能会有优化升级。请以实际标签信息为准。
备注：
以上数据均来自公开文献, Solarbio暂未进行独立验证, 仅供参考。
These protocols are for reference only. Solarbio does not independently validate these
methods.
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Aiping Wang, Na Liu, Jianguo Zhao, Yan Niu, Yumei Chen, Jingming Zhou, Enping Liu, Gaiping Zhang
IF：
3.1000
Publish_to：
JOURNAL OF VIROLOGICAL METHODS
PMID：
38154579
Immunochromatographic test strip for quantitative and rapid detection of tolfenpyrad in food samples
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Author：
Yuan Cheng, Lingling Guo, Aihong Wu, Xinxin Xu, Liqiang Liu, Chuanlai Xu, Hua Kuang, Liguang Xu
IF：
3.0000
Publish_to：
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
PMID：
37524012
Synergism between calcium nitrate applications and fungal endophytes to increase sugar concentration in Festuca sinensis under cold stress
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Author：
Lianyu Zhou, Chunjie Li, James F. White, Richard D. Johnson
IF：
2.9840
Publish_to：
PeerJ
PMID：
35070512
The effect of composite enzyme catalysis whey protein cross-linking on filtration performance
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Author：
Wang Wen-qiong, Zhou Ji-yang, Yu Qian, Li Jianju
IF：
2.8630
Publish_to：
Food Science & Nutrition
PMID：
34136173
Selenium deficiency modulates necroptosis-mediated intestinal inflammation in broiler through the lncRNAWSF27/miRNA1696/GPX3 axis
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Author：
Zhang Yiming, Wei Chunyu, Ding Jiayi, Chu Jiahong, Huang Bo, Shi Guangliang, Li Shu
IF：
2.7000
Publish_to：
JOURNAL OF ANIMAL SCIENCE
PMID：
39331000
Phenolic Compounds of Propolis Alleviate Lipid Metabolism Disorder
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Author：
Lingjie Kong, Yuhao Zhang, Zhouxu Feng, Jie Dong, Hongcheng Zhang
IF：
2.6300
Publish_to：
Evidence-based Complementary and Alternative Medicine
PMID：
33688365
Distinct different expression of Th17 and Th9 cells in coxsackie virus B3-induced mice viral myocarditis
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Author：
Kong Qing, Wu Weifeng, Yang Fan, Yan Yuluan, Pang
IF：
2.4650
Publish_to：
Virology Journal
PMID：
21635745
BPIFA1 alleviates allergic rhinitis by regulating the NF-κB signaling pathway and Treg/Th17 balance
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Author：
Ying Yang, Shidong Li, Hongyan Xu
IF：
2.4000
Publish_to：
International Journal of Rheumatic Diseases
PMID：
39450979
Leucine regulates -amylase and trypsin synthesis in dairy calf pancreatic tissue in vitro via the mammalian target of rapamycin signalling pathway
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Author：
L. Guo (a1), J. H. Yao (a1), C. Zheng (a1), H. B.
IF：
1.8700
Publish_to：
animal
PMID：
30616697
技术问答
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请问用什么溶剂溶解?2023-08-24 15:24:05
您好，可以用水溶解，参考溶解性为10mg/ml。