Malachite Green Phosphate Detection Kit

高靈敏度Malachite green磷酸根呈色法 |貨號303-0125（靈敏度4－40uM）

PiColorLock Gold Phosphate Detection System

PiColorLock™磷酸根檢測試劑，其原理為檢測無機磷酸根（Pi）之含量；使用Malachite green磷酸根呈色法。可用於測量會產生Pi的任何酶，包括ATPases、GTPases, phosphatases、Kinase、Nucleotidases、helicases, 5’nucleotidases, DNA gyrases, heat shock proteins (HSPs), P-type ATPases, Kinesins, transferases and ATP-dependent ligases等酵素活性之分析。PiColorLock™Gold基於malachite green-based assay孔雀石綠的測定方法，用於測量 free phosphate無機磷酸鹽水溶液。 該測定依賴於孔雀石綠之間形成複合物鉬酸鹽和free orthophosphate游離正磷酸鹽，溶液顏色從orange橙色變為綠色 green。顏色的形成可以方便地在分光光度計（620-650nm）上或在分光光度計上測量，讀值與游離正磷酸鹽濃度直接相關。 PiColorLock™的獨特配方提高了測定線性，動態範圍和顏色穩定性；也可以用PiColorLock與不穩定的底物（例如ATP，GTP）一起使用，所述底物與其它酸性染料基檢測試劑產生高的非酶背景信號。此產品為不具放射性應用Malachite green磷酸根呈色法，穩定的產生綠色dye-phosphate複合物，Pi-dye穩定性複合物於長時間之測量，不易產生反應物沉澱(30分後)問題而影響測量讀值。偵測範圍廣，在蛋白質濃度高下不會抑制顏色之形成；可減少樣品稀釋的必要。另提供Stabilizer在酸可溶性受質下，避免發生受質非酵素水解干擾造成的高背景雜訊。

PiColorLock™ 可用於測量以下游離正磷酸鹽的釋放

1. Phosphatase Assays磷酸酶測定：從peptide，protein蛋白質或 small molecule substrate小分子底物中釋放磷酸鹽
2. Lipase Assays脂肪酶測定：從磷脂中釋放phospholipids磷酸鹽
3. Nucleoside Triphosphatase Assays核苷三磷酸酶測定：從nucleoside triphosphates核苷三磷酸中釋放磷酸鹽（ATP，GTP，TTP，CTP）
4. Quantitation 定量phospholipids磷脂，proteins蛋白質和DNA中的phosphate磷酸鹽
5. Drug Discovery藥物發現：磷酸酶抑制劑的高通量篩選。

 

 

 

 

 

Figure 1. Competitor assays are beset with several problems including reagent precipitation. The high stability of PiColorLock™ ensures high stability of the colored dye–phosphate complexes (green color).圖1.市售商品存在試劑沉澱問題。 PiColorLock™的高穩定性確保了有色染料 – 磷酸鹽配合物的高穩定性（綠色）。

 

 

 

 

 

 

 

 

Figure 2. ATP has been incubated in three detection reagents. A steadily rising background signal is seen with competitor reagents, whereas PiColorLock™ provides baseline readings.圖2. ATP在三種檢測試劑。 使用市售商品可以看到穩定上升的背景信號，而PiColorLock™則提供基線讀數。

 

 

 

 

 

 

 

 

 

 

 

Figure 3. PiColorLock™ has also been designed to have a large linear range, thus reducing the need for sample dilution. Competitors’ products are linear over a much narrower range of concentrations.圖3. PiColorLock™具有較大的線性範圍，因此減少了樣品稀釋的需要。市售商品的產品在更窄的濃度範圍內才是線性的。

產品型號	產品名稱	包裝
303-0125 303-0500	PiColorLock Gold Phosphate Detection System（靈敏度4－40uM）	最多1560 assays 最多6250 assays
關聯商品
601-9999	10mM Lyophilized ATP	0.5ml
602-0120 602-0121 602-0122	High Throughput Colorimetric GTPase Assays	196 assays 480 assays 5 × 480 assays
501-0015	PiBind resin－快速移除樣品中污染的磷酸根（Pi）	5g

引用文獻

1. Selective Inhibitors of Alpha2-containing Isoforms of Na,K-ATPase and use thereof for reduction of intraocular pressure (FreePatentsOnline, 2018)
2. Mycobacterial PknG Targets the Rab7l1 Signaling Pathway To Inhibit Phagosome–Lysosome Fusion (Journal of Immunology, 2018)
3. c-Abl-mediated Drp1 phosphorylation promotes oxidative stress-induced mitochondrial fragmentation and neuronal cell death (Nature.com, 2017)
4. Disassembly of the Staphylococcus aureus hibernating 100S ribosome by an evolutionarily conserved GTPase – PNAS (2017)
5. Construction of Structural Mimetics of the Thyrotropin Receptor Intracellular Domain – Biophys J. (2016)
6. Development of a one-pot assay for screening and identification of Mur pathway inhibitors in Mycobacterium tuberculosis – Nature (2016)
7. Selective assembly of Na,K-ATPase ?2?2 heterodimers in the heart: distinct functional properties and isoform-selective inhibitors – J Biol Chem (2016)
8. Reduced Dynamin-related Protein 1 Protects Against Phosphorylated Tau-induced Mitochondrial Dysfunction and Synaptic Damage in Alzheimer’s Disease – Oxford Journals (2016)
9. Structure of the Dictyostelium Myosin-II Heavy Chain Kinase A (MHCK-A) [alpha]-kinase domain apoenzyme reveals a novel autoinhibited conformation – Scientific Rep. (2016)
10. Independent mechanisms target SMCHD1 to H3K9me3-modified chromatin and the inactive X chromosome – Molecular and Cellular Biology (2015)
11. Characterization of the Catalytic and Nucleotide Binding Properties of the Alpha-kinase domain of Dictyostelium Myosin-II Heavy Chain Kinase A. – The Journal of Biological Chemistry (2015)
12. Concerted and differential actions of two enzymatic domains underlie Rad5 contributions to DNA damage tolerance – Nucleic Acids Research (2015)
13. Embryo defective 14 encodes a plastid-targeted cGTPase essential for embryogenesis in maize – Plant J. (2015)
14. Insight into Structure-Function Relationships and Inhibition of the Fatty Acyl-AMP Ligase (FadD32) Orthologs from Mycobacteria – J. Biol. Chem. (2016)
15. Structure-Based Mutagenesis of Sulfolobus Turreted Icosahedral Virus B204 Reveals Essential Residues in the Virion-Associated DNA-Packaging ATPase – J. Virol. (2016)
16. Benzimidazole covalent probes and the gastric H+/K+-ATPase as a model system for protein labeling in a copper-free setting – Molecular BioSystems (2016)
17. Purification of the Human ?2 Isoform of Na,K-ATPase Expressed in Pichia pastoris. Stabilization by Lipids and FXYD1 – Biochemistry (2007)
18. Stabilization of Na+,K+-ATPase Purified from Pichia pastoris Membranes by Specific Interactions with Lipids – Biochemistry (2007)
19. Characterization of Human GTPBP3, a GTP-Binding Protein Involved in Mitochondrial tRNA Modification – Mol Cell Biol (2008)
20. Characterization of Streptococcus gordonii SecA2 as a Paralogue of SecA – J Bacteriol (2009)
21. Structural and Mutational Analyses of Deinococcus radiodurans UvrA2 Provide Insight into DNA Binding and Damage Recognition by UvrAs – Structure (2009)
22. Identification of a Small Molecule Inhibitor of the Human DNA Repair Enzyme Polynucleotide Kinase/Phosphatase – Cancer Res (2009)
23. Functional study on GTP hydrolysis by the GTP-binding protein from Sulfolobus solfataricus, a member of the HflX family – J Biochem (2010)
24. Oligomeric assembly and interactions within the human RuvB-like RuvBL1 and RuvBL2 complexes – Biochem. J. (2010)
25. Anti-tubercular screening of natural products from Colombian plants: 3-methoxynordomesticine, an inhibitor of MurE ligase of Mycobacterium tuberculosis – J Antimicrob Chemother (2010)
26. Roles of Electrostatics and Conformation in Protein-Crystal Interactions – PLoS One (2010)
27. Sensor combination and chemometric variable selection for online monitoring of Streptomyces coelicolor fed-batch cultivations – Applied Microbiology and Biotechnology (2010)
28. Mechanism of Action of an Imidopiperidine Inhibitor of Human Polynucleotide Kinase/Phosphatase – JBC (2010)
29. ATP-dependent MurE ligase in Mycobacterium tuberculosis: Biochemical and structural characterisation – Tuberculosis (2010)
30. Selectivity of Digitalis Glycosides for Isoforms of Human Na,K-ATPase – JBC (2010)
31. Substrate uptake, phosphorus repression, and effect of seed culture on glycopeptide antibiotic production: Process model development and experimental validation – Biotechnology and Bioengineering (2010)
32. Characterisation of multiple substrate-specific (d)ITP/(d)XTPase and modelling of deaminated purine nucleotide metabolism – MIMS EPrints (2011
33. Stabilization of the ?2 isoform of Na,K-ATPase by mutations in a phospholipid binding pocket – JBC (2011)
34. Structural and functional insights into DNA-end processing by the archaeal HerA helicase–NurA nuclease complex – Nucl Acids Res (2011)
35. FXYD Proteins Stabilize Na,K-ATPase, Amplification Of Specific Phosphatidylserine -Protein Interactions – JBC (2011)
36. Sepsis impairs alveolar epithelial function by downregulating Na-K-ATPase pump – AJP – Lung Physiol (2011)
37. Structural and functional insights into DNA-end processing by the archaeal HerA helicase–NurA nuclease complex – Nucleic Acids Research (2011)
38. Sepsis impairs alveolar epithelial function by downregulating Na-K-ATPase pump – AJP – Lung Physiol (2011)
39. Synthesis and Antibacterial Evaluation of a New Series of N-Alkyl-2-alkynyl/(E)-alkenyl-4-(1H)-quinolones – Molecules (2012)
40. The unusual mycobacterial chaperonins: evidence for in vivo oligomerization and specialization of function – Molecular Microbiology (2012)
41. Kinetic Modelling of GlmU Reactions – Prioritization of Reaction for Therapeutic Application – PLoS One (2012)
42. Mesenchymal Stem Cell Therapy – Humana Press (2013)
43. Structural and Mechanistic Characterization of L-Histidinol Phosphate Phosphatase from the PHP Family of Proteins – Biochemistry (2013)
44. Polyphosphates inhibit extracellular matrix mineralization in MC3T3-E1 osteoblast cultures – Bone (2013)
45. A Newly Uncovered Group of Distantly Related Lysine Methyltransferases Preferentially Interact with Molecular Chaperones to Regulate Their Activity – PLoS Genetics (2013)
46. Na+ Regulation in the Malaria Parasite Plasmodiumfalciparum Involves the Cation ATPase PfATP4 and Is a Target of the Spiroindolone Antimalarials – Cell Host & Microbe (2013)
47. A calcineurin docking motif (LxVP) in dynamin-related protein 1 contributes to mitochondrial fragmentation and ischemic neuronal injury – The Journal of Biological Chemistry (2013)
48. Characterisation of ATP-Dependent Mur Ligases Involved in the Biogenesis of Cell Wall Peptidoglycan in Mycobacterium tuberculosis – PLoS One (2013)
49. ATP-dependent transport of statins by human and rat MRP2/Mrp2 – Toxicology and Applied Pharmacology (2013)
50. Structural and functional characterization of a phosphatase domain within yeast general transcription factor TFIIIC – The Journal of Biological Chemistry (2013)
51. Plastid division control: the PDV proteins regulate DRP5B dynamin activity – Plant Molecular Biology (2013)
52. Extracellular ATP Released by Osteoblasts Is A Key Local Inhibitor of Bone Mineralisation – PLoS One (2013)
53. Direct regulation of microtubule dynamics by KIF17 motor and tail domains – Journal of Biological Chemistry (2013)
54. A novel cholesterol-producing Pichia pastoris strain is an ideal host for functional expression of human Na,K-ATPase ?3?1 isoform – Applied Microbiology and Biotechnology (2013)
55. Chrysophaentins are competitive inhibitors of FtsZ and inhibit Z-ring formation in live bacteria – Bioorganic & Medicinal Chemistry (2013)
56. RNA silencing of genes involved in Alzheimer’s disease enhances mitochondrial function and synaptic activity – Biochimica et Biophysica Acta (2013)
57. A biosensor of local kinesin activity reveals roles of PKC and EB1 in KIF17 activation – Journal of Cell Biology (2013)
58. 3,4-Methylenedioxy-?-nitrostyrene inhibits NLRP3 activation by blocking assembly of the inflammasome – Journal of Biological Chemistry (2013)
59. Bordetella pertussis adenylate cyclase toxin translocation across a tethered lipid bilayer – PNAS (2013)
60. Mutations in the Chromodomain-like Insertion of Translation Elongation Factor 3 Compromise Protein Synthesis Through Reduced ATPase Activity – Journal of Biological Chemistry (2013)
61. Prospecting for Unannotated Enzymes: Discovery of a 3?,5?-Nucleotide Bisphosphate Phosphatase within the Amidohydrolase Superfamily – Biochemistry (2013)
62. Ablation of Osteopontin Improves the Skeletal Phenotype of Phospho1-/- Mice – Journal of Bone and Mineral Research (2014)
63. Low Molecular Weight Protein Tyrosine Phosphatases Control Antibiotic Production in Streptomyces coelicolor A3(2) – Enzyme Engineering (2014)
64. The PIKE Homolog Centaurin gamma Regulates Developmental Timing in Drosophila – PLoS One (2014)
65. Quantification of biliary excretion and sinusoidal excretion of 5(6)-carboxy-2?,7?-dichlorofluorescein (CDF) in cultured hepatocytes isolated from Sprague Dawley, Wistar and Mrp2-deficient Wistar (TR?) rats – Toxicology in Vitro (2014)
66. Self-Assembly of MinE on the Membrane Underlies Formation of the MinE-Ring to Sustain Function of the E. coli Min System – Journal of Biological Chemistry (2014)
67. Allosteric Activation of Bordetella pertussis Adenylyl Cyclase by Calmodulin: Molecular Dynamics and Mutagenesis Studies – Journal of Biological Chemistry (2014)
68. Optimization of Pyrrolamides as Mycobacterial GyrB ATPase Inhibitors: Structure-Activity Relationship and In Vivo Efficacy in a Mouse Model of Tuberculosis – Antimicrobial Agents and Chemotherapy (2014)
69. Assays, Surrogates, and Alternative Technologies for a TB Lead Identification Program Targeting DNA Gyrase ATPase – Journal of Biomolecular Screening (2014)