Alkaline Phosphatase Assay Kit 碱性磷酸酶活性检测试剂盒(显色法)

【温馨提示I】:见碱性磷酸酶产品专题,了解更多相关产品资料;

【温馨提示II】:见碱性磷酸酶染色试剂盒产品

【温馨提示III】:见碱性磷酸酶常用生化底物及试剂盒产品;

产品关键词:

Phosphatase, Alkaline碱性磷酸酶;ALP/AKP;对硝基苯酚磷酸盐(pNPP);偶氮偶联法;显色法(定量);染色法(定性);

基本介绍:

碱性磷酸酶(Phosphatase, AlkalineALP/AP/AKP/ALKP/ALPaseEC 3.1.3.1),是一种含两个相同亚基的二聚体,也是一种约含12%碳水化合物(6%己糖和6%其他中性糖)的糖蛋白,每个ALP分子含4个锌离子和4个二硫键。镁离子存在情况下,ALP可达到最高酶活。碱性条件下,ALP能催化磷酸单酯水解,比如对硝基苯酚磷酸盐(pNPP)、磷酸苯酯、葡萄糖-6-磷酸、葡萄糖-1-磷酸、果糖-6-磷酸、腺苷-5-磷酸等。

常见的碱性磷酸酶ALP包括肠道碱性磷酸酶(如小牛肠来源,calf intestinal)、胎盘碱性磷酸酶(placental alkaline phosphatase)、非组织特异性碱性磷酸酶(tissue-nonspecific isozyme, alkaline phosphatase)等。

ALP常用来偶联抗体和其他蛋白,从而满足ELISAWB和组织化学检测上的应用。通常用来蛋白的去磷酸化实验,比如酪蛋白和核酸等。当需要高检测灵敏度时也能用来蛋白标记。还可用在DNARNA5′-末端去磷酸以防止自连。经碱性磷酸酶的去磷酸化作用后,DNARNA还可被放射性标记磷酸盐标记(通过T4多核苷酸激酶)。

研究意义:

碱性磷酸酶(ALP)活性的变化与一系列生理和病理事件紧密关联,比如骨骼发育、骨骼相关疾病、妊娠相关疾病和炎症性肠病(inflammatory bowel diseases)等。正因如此,ALP被用作衡量生理特征或疾病的参照指标。另外,ALP活性检测在环境监测、食品质量控制等方面也呈现重要作用。

干细胞如诱导多潜能干细胞iPSALP活性很高,则ALP常被用作iPS诱导成功的标志;

分化的结肠癌细胞内ALP活性显著提高,则ALP常被当做结肠癌细胞分化程度的指标(定性/定量);

血清中ALP水平提高,被称作高碱性磷酸酶血症,此症状被认为和恶性胆管阻塞、原发性胆管硬化、原发性硬化胆管炎、肝淋巴瘤和肝肉瘤等肝胆疾病密切相关;

ALP是成骨细胞的副产物,血清中ALP活性升高和骨头发育密切关联;

血清ALP活性过低也与一些疾病相关;

儿童和孕妇血清内ALP活性较常人高一些;血清中ALP活性范围在20-140U/L

检测原理(碱性磷酸酶活性):

对硝基苯酚磷酸盐(pNPP)是一种常用的碱性磷酸酶(ALP)显色底物,碱性条件下能被ALP催化生成对硝基酚(para-nitrophenolp-nitrophenol),呈黄色产物,可在400-415nm检测到吸光度。产物黄色越深,代表ALP活性越高,反之则酶活性越低。则通过比色分析可定量检测ALP水平。

反应式如下:

检测对象:

细胞或组织裂解液或匀浆液、血清、血浆、尿液等样品内源性碱性磷酸酶活性。

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品牌 名称 货号 规格 目录价(元)
Anaspec SensoLyte® pNPP Alkaline Phosphatase Assay Kit *Colorimetric* AS-72146 500 assays 2422

试剂盒组分:

Component Description Quantity
Component A pNPP, colorimetric alkaline phosphatase substrate 25 mL
Component B 10X Assay buffer 50 mL
Component C Stop solution 25 mL
Component D Triton-X-100 500μl
Component E Alkaline Phosphatase Standard, Calf Intestine 10μg /mL, 50μl

AS-72146 SensoLyte pNPP Kit Datasheet

订购信息II国产碱性磷酸酶检测试剂盒,在保证优秀质量的情况下,提供更有竞争力的价格,欢迎选购。

品牌 名称 货号 规格 目录价(元)
MKbio Alkaline Phosphatase Assay Kit (Colorimetric)碱性磷酸酶检测试剂盒(显 色法) MP7501-100T 100T 495

试剂盒组分:

Component Description Quantity
Component A pNPP显色底物 2
Component B 检测缓冲液 15 mL
Component C 终止液 12 mL
Component D 对硝基苯酚溶液(10mM 100μl

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