【溫馨提示II】：見(jiàn)堿性磷酸酶染色試劑盒產(chǎn)品；

【溫馨提示III】：見(jiàn)堿性磷酸酶常用生化底物及試劑盒產(chǎn)品；

產(chǎn)品關(guān)鍵詞：

Phosphatase, Alkaline堿性磷酸酶；ALP/AKP；對硝基苯酚磷酸鹽（pNPP）；偶氮偶聯(lián)法；顯色法（定量）；染色法（定性）；

基本介紹：

? 堿性磷酸酶（Phosphatase, Alkaline，ALP/AP/AKP/ALKP/ALPase，EC 3.1.3.1），是一種含兩個(gè)相同亞基的二聚體，也是一種約含12%碳水化合物（6%己糖和6%其他中性糖）的糖蛋白，每個(gè)ALP分子含4個(gè)鋅離子和4個(gè)二硫鍵。鎂離子存在情況下，ALP可達到最高酶活。堿性條件下，ALP能催化磷酸單酯水解，比如對硝基苯酚磷酸鹽（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。

? 常見(jiàn)的堿性磷酸酶ALP包括腸道堿性磷酸酶（如小牛腸來(lái)源，calf intestinal）、胎盤(pán)堿性磷酸酶（placental alkaline phosphatase）、非組織特異性堿性磷酸酶（tissue-nonspecific isozyme, alkaline phosphatase）等。

? ALP常用來(lái)偶聯(lián)抗體和其他蛋白，從而滿(mǎn)足ELISA、WB和組織化學(xué)檢測上的應用。通常用來(lái)蛋白的去磷酸化實(shí)驗，比如酪蛋白和核酸等。當需要高檢測靈敏度時(shí)也能用來(lái)蛋白標記。還可用在DNA或RNA 的5′-末端去磷酸以防止自連。經(jīng)堿性磷酸酶的去磷酸化作用后，DNA 或 RNA 還可被放射性標記磷酸鹽標記（通過(guò) T4 多核苷酸激酶）。

研究意義：

堿性磷酸酶（ALP）活性的變化與一系列生理和病理事件緊密關(guān)聯(lián)，比如骨骼發(fā)育、骨骼相關(guān)疾病、妊娠相關(guān)疾病和炎癥性腸?。?/span>inflammatory bowel diseases）等。正因如此，ALP被用作衡量生理特征或疾病的參照指標。另外，ALP活性檢測在環(huán)境監測、食品質(zhì)量控制等方面也呈現重要作用。

? 干細胞如誘導多潛能干細胞iPS，ALP活性很高，則ALP常被用作iPS誘導成功的標志；

? 分化的結腸癌細胞內ALP活性顯著(zhù)提高，則ALP常被當做結腸癌細胞分化程度的指標（定性/定量）；

? 血清中ALP水平提高，被稱(chēng)作高堿性磷酸酶血癥，此癥狀被認為和惡性膽管阻塞、原發(fā)性膽管硬化、原發(fā)性硬化膽管炎、肝淋巴瘤和肝肉瘤等肝膽疾病密切相關(guān)；

? ALP是成骨細胞的副產(chǎn)物，血清中ALP活性升高和骨頭發(fā)育密切關(guān)聯(lián)；

? 血清ALP活性過(guò)低也與一些疾病相關(guān)；

? 兒童和孕婦血清內ALP活性較常人高一些；血清中ALP活性范圍在20-140U/L。

檢測原理（堿性磷酸酶活性）：

對硝基苯酚磷酸鹽（pNPP）是一種常用的堿性磷酸酶（ALP）顯色底物，堿性條件下能被ALP催化生成對硝基酚（para-nitrophenol，p-nitrophenol），呈黃色產(chǎn)物，可在400-415nm檢測到吸光度。產(chǎn)物黃色越深，代表ALP活性越高，反之則酶活性越低。則通過(guò)比色分析可定量檢測ALP水平。

反應式如下：

檢測對象：

細胞或組織裂解液或勻漿液、血清、血漿、尿液等樣品內源性堿性磷酸酶活性。

訂購信息I：Anaspec國原裝進(jìn)口，>幾十篇文獻引用，高口碑產(chǎn)品。

試劑盒組分：

AS-72146 SensoLyte pNPP Kit Datasheet

訂購信息II：國產(chǎn)堿性磷酸酶檢測試劑盒，在保證優(yōu)秀質(zhì)量的情況下，提供更有競爭力的價(jià)格，歡迎選購。

試劑盒組分：

引用文獻（Anaspec，AS-72146）：

Knopp, M. et al. (2015). Amelioration of the fitness costs of antibiotic resistance due to reduced outer membrane permeability by upregulation of alternative porins MolBioEvol 32doi:10.1093/molbev/msv195

Dai, J. et al. (2014). Cabozantinib Inhibits Prostate Cancer Growth and Prevents Tumor-Induced Bone Lesions. Clin Cancer Res 20, 617. doi: 10.1158/1078-0432.CCR-13-0839.

Okon, IS. et al. (2014). Liver kinase B1 expression promotes phosphatase activity and abrogation of receptor tyrosine kinase phosphorylation in human cancer cells. J Biol Chem 289, 1639.

Mah, YJ. et al. (2014). The effect of epigallocatechin-3-gallate (EGCG) on human alveolar bone cells both in vitro and in vivo. Archives Oral Biol 59, 539.

Pauksch, L. et al. (2014). Biocompatibility of silver nanoparticles and silver ions in primary human mesenchymal stem cells and osteoblasts. Acta Biomat 10, 439.

Rebollo, A. et al. (2014). Liquid fructose downregulates Sirt1 expression and activity and impairs the oxidation of fatty acids in rat and human liver cells. Biochim Biophys Acta -Mol Cell Biol Lipids1841, 514.

Yamano, S. et al. (2014). The effect of a bioactive collagen membrane releasing PDGF or GDF-5 on bone regeneration. Biomat 35, 2446.

Choi, B. et al. (2012). Facile surface functionalization with glycosaminoglycans by direct coating with mussel adhesive protein. Tissue Eng Part C Meth 18, 71.

Kamiyama, M. et al. (2012). The establishment of a primary culture system of proximal tubule segments using specific markers from normal mouse kidneys. Intl J Mol Sci 13, 5098. doi:10.3390/ijms13045098

Kim, S-M. et al. (2012).Gelatin-layered and multi-sized porous β-tricalcium phosphate for tissue engineering scaffold. Nanoscale Res Lett 7, 78. doi:10.1186/1556-276X-7-78Open Access

Martinez-Conesa, E. et al. (2012). Characterization of ocular surface epithelial and progenitor cell markers in human adipose stromal cells derived from lipoaspirates. Invest. Ophthalmol. Vis. Sci 53, 513-520.

Sola, A. et al. (2012). Heat treatment of Na2O-CaO-P2O5-SiO2 bioactive glasses: Densification processes and postsintering bioactivity. J Biomed Mat Res 100A, 305. doi:10.1002/jbm.a.33276

Sun, J. et al. (2012). Biomimetic interpenetrating polymer network hydrogels based on methacrylated alginate and collagen for 3D pre-osteoblast spreading and osteogenic differentiation. Soft Matter 8, 2398.

Sunada, K. et al. (2012). Highly efficient antiviral and antibacterial activities of solid-state cuprous compounds. J Haz Mat 235-236, 265. doi:10.1002/jbm.a.33276

Thorpe, M. et al. (2012). The external mechanical environment can override the influence of local substrate in determining stem cell fate. J Biomechanics 45, 2483.

Vinardell, T. et al. (2012). Hydrostatic pressure acts to stabilize a chondrogenic phenotype in porcine joint tissue derived stem cells. Euro Cells and Mat 23, 121.

Williams, C. et al. (2012). A comparison of human smooth muscle and mesenchymal stem cells as potential cell sources for tissue-engineered vascular patches. Tissue Eng Part A.

Abedini, S. et al. (2011). Effects of cryopreservation with a newly-developed magnetic field programmed freezer on periodontal ligament cells and pulp tissues. Cryobiol10.1016/j.cryobiol.2011.03.001.

Brammer, K. et al. (2011). Highly bioactive 8 nm hydrothermal TiO2 nanotubes elicit enhanced bone cell response. Adv Eng Mat 13, B88.

Brammer, K. et al. (2011). Comparative cell behavior on carbon-coated TiO2 nanotube surfaces for osteoblasts vs. osteo-progenitor cells. Acta Biomaterialia 7, 2697.

Frandsen, C. et al. (2011). Zirconium oxide nanotube surface prompts increased osteoblast functionality and mineralization. Mat Sci and Eng 31, 1716.

Guller, S. et al. (2011). Protein composition of microparticles shed from human placenta during placental perfusion: Potential role in angiogenesis and fibrinolysis in preeclampsia. Placenta 32, 63.

Hong, D. et al. (2011). Quantitative proteomic analysis of dexamethasone-induced effects on osteoblast differentiation, proliferation, and apoptosis in MC3T3-E1 cells using SILAC. Osteo Inter22, 2175.

Jeon, O. et al. (2011). Affinity-based growth factor delivery using biodegradable, photocrosslinked heparin-alginate hydrogels. J of Cont Rel 154, 258.

Kemoun, P. et al. (2011). The role of cell surface markers and enamel matrix derivatives on human periodontal ligament mesenchymal progenitor responses in vitro. Biomat 32, 7375.

Kulp, A. et al. (2011). Recognition of β-Strand motifs by RseB is required for E activity in escherichia coli. J. Bacteriol 193, 6179-6186.

Lau, E. et al. (2011). Effect of low-magnitude, high-frequency vibration on osteogenic differentiation of rat mesenchymal stromal cells. J of Ortho Res 29, 1075.

Lin, F. et al. (2011). Role of mitogen-activated protein kinase in osteoblast differentiation. J Ortho Res 29, 204.

Manning, A. et al. (2011). Contribution of bacterial outer membrane vesicles to innate bacterial defense Biomedcentral 11, 258.

McNeil, S. et al. (2011). Polycaprolactone fibres as a potential delivery system for collagen to support bone regeneration. Curr Drug Deliv 8, 448.

Nazari-Shafti, T. et al. (2011). Mesenchymal stem cell derived hematopoietic cells are permissive to HIV-1 infection. Retrovirol 8, 3.

Oh, P-S. et al. (2011). Schlafen-3 decreases cancer stem cell marker expression and autocrine/juxtacrine signaling in FOLFOX-resistant colon cancer cells. AJP-GI 10.1152/ajpgi.00403.

Rathbone, C. et al. (2011). Effect of various concentrations of antibiotics on osteogenic cell viability and activity. J Orthopaedic Res 10.1002/jor.21343.

Salerno, A. et al. (2011). Processing/structure/property relationship of multi-scaled PCL and PCL–HA composite scaffolds prepared via gas foaming and NaCl reverse templating. Biotech and Bioengin 108, 963.

Shah, D. et al. (2011). Cell invasion of poultry-associated Salmonella Enteritidis isolates is associated with pathogenicity, motility and secretion of type-three secretion system secreted proteins. Microbiol 10.1099/mic.0.044461-0.

Tan, F. et al. (2011). Osteoconductivity and growth factor production by MG63 osteoblastic cells on bioglass-coated orthopedic implants. Biotech and Bioengin 108, 454.

Uhrig, R. et al. (2011). Two ancient bacterial-like PPP family phosphatases from arabidopsis are highly conserved plant proteins that possess unique properties. Plant Physiology 157, 1778-1792.

Wang, J. et al. (2011). The effect of scaffold architecture on odontogenic differentiation of human dental pulp stem cells. Biomaterials 32, 7822.

Yamano, S. et al. (2011). Bioactive collagen membrane as a carrier for sustained release of PDGF.J Tis Sci Eng 2, 4.

Zhou, Q. et al. (2011). Localized colonic stem cell transplantation enhances tissue regeneration in murine colitis. J Cell Mol Med

Barbier de La Serre, C. et al. (2010). Propensity to high-fat diet-induced obesity in rats is associated with changes in the gut microbiota and gut inflammation. Am J Physiol Gastrointest Liver Physiol 299, G440.

Choi, B. et al. (2010). Cell behavior on extracellular matrix mimic materials based on mussel adhesive protein fused with functional peptides. Biomaterials 31, 8980.

Cramer, C. et al. (2010). Persistent high glucose concentrations alter the regenerative potential of mesenchymal stem cells. Stem Cells Dev 19, 1875.

Fan, Q. et al. (2010). Requirement of TGFβ signaling for SMO-mediated carcinogenesis. J Biol Chem285, 36570.

Hildreth, B. et al. (2010). PTHrP 1-141 and 1-86 increase in vitro bone formation. J Surg Res 162, e9.

Hong, D. et al. (2010). Morphological and proteomic analysis of early stage of osteoblast differentiation in osteoblastic progenitor cells. Experimental Res 316, 2291.

Hu, J. et al. (2010). Response of human embryonic stem cells derived mesenchymal stem cells to osteogenic factors and architectures of materials during in vitro osteogenesis. Tissue Engineer Part A 10.1089/ten.TEA.2010.0097.

Kanwar, S. et al. (2010). The Wnt/beta-catenin pathway regulates growth and maintenance of colonospheres. Mol Cancer 6, 212.

Kemmis, C. et al. (2010). Bone morphogenetic protein 6 drives both osteogenesis and chondrogenesis in murine adipose-derived mesenchymal cells depending on culture conditions.Biochem Biophys Res Commun. 401, 20.

Lu, C. et al. (2010). MT1-MMP controls human mesenchymal stem cell trafficking and differentiation. Blood 115, 221.

Misra, S. et al. (2010). Effect of nanoparticulate bioactive glass particles on bioactivity and cytocompatibility of poly(3-hydroxybutyrate) composites. J R Social Interface 7, 453.

Oku, S. et al. (2010). JAK2 V617F uses distinct signalling pathways to induce cell proliferation and neutrophil activation. British journal of Haematology 150, 334.

Raucci, MG. et al. (2010). Biomineralized porous composite scaffolds prepared by chemical synthesis for bone tissue regeneration. Acta Biomater 10.1016/j.actbio.2010.04.018.

Thi, MM. et al. (2010). Characterization of hTERT-immortalized osteoblast cell lines generated from wild-type and connexin43-null mouse calvaria. Am J Physiol Cell Physiol 299, C994.

Wang, J. et al. (2010). The odontogenic differentiation of human dental pulp stem cells on nanofibrous poly(l-lactic acid) scaffolds in vitro and in vivo. Acta Biomaterialia 6, 3856.

Zhang, Z. et al. (2010). A comparison of bioreactors for culture of fetal mesenchymal stem cells for bone tissue engineering. Biomaterials 31, 8684.

Hemauer, S. et al. (2009). Opiates inhibit paclitaxel uptake by P-glycoprotein in preparations of human placental inside-out vesicles. Biochem Pharm 78, 1272.

Kurimoto-Nakano, R. et al. (2009). Replicative senescence of vascular smooth muscle cells enhances the calcification through initiating the osteoblastic transition. Am J Physiol Heart Circ Physiol 297, H1673-H1684

Lee, J. et al. (2009). Modular peptides promote human mesenchymal stem cell differentiation on biomaterial surfaces. Acta Biomater 6, 21.

Pennisi, A. et al. (2009). Inhibitor of DASH proteases affects expression of adhesion molecules in osteoclasts and reduces myeloma growth and bone disease. British J of Hematology 145, 775.

Stanisic, V. et al. (2009). OTU domain-containing ubiquitin aldehyde-binding protein 1 (OTUB1) deubiquitinates estrogen receptor (ER) α and affects ERα transcriptional activity. J Biol Chem 284, 16135.

Zhang, Z. et al. (2009). Superior osteogenic capacity for bone tissue engineering of fetal compared with perinatal and adult mesenchymal stem cells. Stem Cells 27, 126.

Haraguchi, N. et al. (2008). CD133+CD44+ population efficiently enriches colon cancer initiating cells. Ann Surg Oncol 15, 2927.

Meng, Q. et al. (2008). Regulating the age-related oxidative damage, mitochondrial integrity, and antioxidative enzyme activity in fischer 344 rats by supplementation of the antioxidant epigallocatechin-3-gallate. Rejuv Res 11, 649.

Scaffidi, P. et al. (2008). Lamin A-dependent misregulation of adult stem cells associated with accelerated ageing. Nature Cell Biol 10, 452.

Bae, J. et al. (2007). Engineering a disulfide bond to stabilize the calcium binding loop of activated protein C eliminates its anticoagulant but not protective signaling properties. J Biol Chem10.1074/jbc.M610547200.

Yang, L. et al. (2007). Identification of a specific exosite on activated protein C for interaction with protease-activated receptor 1. J Biol Chem 282, 25493.

 — —Written/Edited by V. Shallan【版權歸MKBio懋康所有】