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473:
PNPP is classified as a chromogenic substrate because of its ability to transform from a colorless compound to a colored compound through a biological mechanism, dephosphorylation. PNPP is used because of its low cost and the rate of the reactions can be measured over a wide range of substrate
477:
A PNPP assay involves mixing the sample with a PNPP-containing mixture and permitting the reaction to run its course for a predetermined period of time. After that point, the process is halted through the addition of a stop solution, often made of a potent alkali like sodium hydroxide.
474:
concentrations because the concentration of the substrate is not a limiting factor in the reaction. The limitations of PNPP is that it is a small molecule and perhaps does not entirely represent the conditions and structures that are encountered physiologically.
470:(pNP). The resulting phenolate is yellow, with a maximal absorption at 405 nm. This property can be used to determine the activity of various phosphatases including alkaline phosphatase (AP) and protein tyrosine phosphatase (PTP).
481:
The substance is sensitive to light, and thus should be stored protected from light. This is also important after adding the substrate to the mixture and before reading. −20 °C is the optimal storage temperature.
485:
To get precise and credible findings, the assay parameters must be carefully regulated because the reaction is sensitive to factors such as pH, temperature, and the varying degree of inhibitors or stimulants.
422:
751:"Kinetics of the Alkaline Phosphatase Catalyzed Hydrolysis of Disodium p-Nitrophenyl Phosphate: Effects of Carbohydrate Additives, Low Temperature, and Freezing"
325:
118:
791:
621:
National Center for
Biotechnology Information (2023). PubChem Compound Summary for CID 4686862, p-Nitrophenyl phosphate. Retrieved May 14, 2023 from
463:
and conventional spectrophotometric assays. Phosphatases catalyze the hydrolysis of pNPP liberating inorganic phosphate and the conjugate base of
528:
816:
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811:
686:"Enhanced Hydrolysis of p -Nitrophenyl Phosphate by Iron (Hydr)oxide Nanoparticles: Roles of Exposed Facets"
39:
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Li, Tong; Zhong, Wen; Jing, Chuanyong; Li, Xuguang; Zhang, Tong; Jiang, Chuanjia; Chen, Wei (2020-07-21).
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InChI=1S/C6H6NO6P/c8-7(9)5-1-3-6(4-2-5)13-14(10,11)12/h1-4H,(H2,10,11,12)
309:
InChI=1/C6H6NO6P/c8-7(9)5-1-3-6(4-2-5)13-14(10,11)12/h1-4H,(H2,10,11,12)
209:
766:
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Terefe, N.S.; Arimi, J.M.; VanLoey, A.; Hendrickx, M. (2004-10-01).
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Except where otherwise noted, data are given for materials in their
460:
129:
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https://pubchem.ncbi.nlm.nih.gov/compound/p-Nitrophenyl-phosphate
577:"Derivative absorption spectrophotometry of native proteins"
259:
38:
29:
637:"Analysis of Protein Tyrosine Phosphatases and Substrates"
575:
Matsushima, Ayako; Inoue, Yorinao; Shibata, Kazuo (1975).
417:
792:pNPP disodium salt hexahydrate (SigmaAldrich.com)
221:
93:
635:Mercan, Fatih; Bennett, Anton M. (July 2010).
563:Protein Phosphorylation: A Practical Approach
561:MacKintosh, C. (1993). In D.G. Hardie (Ed.).
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15:
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725:"p-Nitrophenyl Phosphate (PNPP) - NEB"
690:Environmental Science & Technology
641:Current Protocols in Molecular Biology
190:
505:"Protein Tyrosine Phosphatase Assays"
302:Key: XZKIHKMTEMTJQX-UHFFFAOYSA-N
157:
137:
7:
312:Key: XZKIHKMTEMTJQX-UHFFFAOYAU
212:
63:4-Nitrophenyl dihydrogen phosphate
14:
407:
509:Current Protocols in Immunology
403:(at 25 °C , 100 kPa).
457:alkaline and acid phosphatases
1:
503:Lorenz, Ulrike (2017-05-07).
653:10.1002/0471142727.mb1816s91
593:10.1016/0003-2697(75)90520-5
521:10.1002/0471142735.im1107s93
565:. 221. New York: IRL Press.
333:C1=CC(=CC=C1(=O))OP(=O)(O)O
25:
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451:) is a non-proteinaceous
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817:Nitrobenzene derivatives
702:10.1021/acs.est.9b07473
581:Analytical Biochemistry
755:Biotechnology Progress
723:Biolabs, New England.
43:
34:
20:-Nitrophenylphosphate
453:chromogenic substrate
445:-Nitrophenylphosphate
42:
33:
376:219.09
59:Preferred IUPAC name
797:Nunc Brand Products
21:
430:Infobox references
381:Related compounds
44:
35:
16:
767:10.1021/bp0498894
696:(14): 8658–8667.
438:Chemical compound
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387:Related compounds
255:CompTox Dashboard
119:Interactive image
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812:Organophosphates
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468:-nitrophenol
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78:Identifiers
70:Other names
17:
729:www.NEB.com
342:Properties
197:100.005.777
159:ChEMBL24231
139:CHEBI:17440
806:Categories
734:29 October
490:References
372:Molar mass
243:59NF9TE3BA
170:ChemSpider
106:3D model (
85:CAS Number
775:8756-7938
710:0013-936X
661:1934-3639
601:0003-2697
549:21462163
459:used in
392:Paraoxon
366:P
95:330-13-2
670:4351734
609:1169006
540:3097125
423:what is
421: (
210:PubChem
773:
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418:verify
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326:SMILES
150:ChEMBL
53:Names
647:(1).
461:ELISA
291:InChI
130:ChEBI
108:JSmol
771:ISSN
736:2017
706:ISSN
657:ISSN
605:PMID
597:ISSN
545:PMID
525:ISBN
466:para
455:for
449:pNPP
443:para
234:UNII
72:pNPP
18:para
763:doi
698:doi
665:PMC
649:doi
589:doi
535:PMC
517:doi
260:EPA
223:378
213:CID
179:369
808::
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362:NO
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447:(
413:N
364:6
360:6
358:H
356:6
354:C
262:)
258:(
110:)
Text is available under the Creative Commons Attribution-ShareAlike License. Additional terms may apply.
