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N1127 Sigma

2-Nitrophenyl β-D-galactopyranoside

≥98% (enzymatic)

Synonym: o-Nitrophenyl β-D-galactopyranoside, ONPG

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Description

General description

ONPG (2-Nitrophenyl β-D-galactopyranoside) is a colorimetric substrate for β-galactosidase.

Packaging

1 g in poly tube

250 mg in poly bottle

5, 25 g in poly bottle

500 mg in poly tube

Principle

β-galactosidase, breaks down lactose into galactose and glucose. β-Galactosidase is not lactose specific and can act on simple galactosides. 2-Nitrophenyl β-D-galactopyranoside hydrolysis results in the release of galactose and a yellow chromogenic compound. The test substrate does not depend on an induced or constitutive permease enzyme to enter the cell, therefore reactions are rapid and occur within a 24-hour period.

Reconstitution

A stock solution can be prepared in molecular biology grade water at a concentration of 3 mg/ml. Alternatively, a stock solution of approximately 20.5 mg/ml is prepared in 100 mM sodium phosphate buffer (pH 7.3). Gentle warming may be required to completely dissolve the product.

Price and Availability


Synthetic Biology Resource Collection

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Safety & Documentation

Safety Information

WGK Germany 
3

Protocols & Articles

Articles

2-Nitrophenyl β-D-galactopyranoside

This product (ONPG) is a colorimetric substrate for β-galactosidase. This product is hydrolyzed to o-nitrophenol by the enzyme. o-Nitrophenol is yellow in alkaline solution and can be quantitated at ...
Keywords: Extinction coefficient, Melting

Protocols

Introduction to Yeast Transformation

Transformation is the process by which exogenous DNA is introduced into a cell, resulting in an inheritable change or genetic modification. This was first reported in Streptococcus pneumoniae by Grif...
Keywords: Cell disruption, Food & Beverage, Gene expression, Genetic, Molecular biology, Pharmaceutical, Polymerase chain reaction, transformation

Yeast Transformation Protocols

Yeasts are considered model systems for eukaryotic studies as they exhibit fast growth and have dispersed cells. Moreover, replica plating and mutant isolation of yeast cells can be done with relativ...
Keywords: Degradations, Genetic, Molecular biology, Transfection, transformation

Peer-Reviewed Papers

References

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Functional Interaction Map of Lyssavirus Phosphoprotein: Identification of the Minimal Transcription Domains Yves Jacob, Eléonore Real, et al. J. Virol. 75, 9613-9622, (2001)

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The DNA-binding domain of CaNdt80p is required to activate CDR1 involved in drug resistance in Candida albicans Jang-Shiun Wang, Yun-Liang Yang, et al. J. Med. Microbiol. 55, 1403-1411, (2006)

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CCAAT/Enhancer Binding Protein β2 Is Involved in Growth Hormone-Regulated Insulin-Like Growth Factor-II Gene Expression in the Liver of Rainbow Trout (Oncorhynchus mykiss) Jay H. Lo and Thomas T. Chen Endocrinology 151, 2128-2139, (2010)

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Dual mechanism of bacterial lethality for a cationic sequence-random copolymer that mimics host-defense antimicrobial peptides. Epand RF, Mowery BP, Lee SE, et al. J. Mol. Biol. 379(1), 38-50, (2008)

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Heterologous expression of glycoside hydrolase family 2 and 42 β-galactosidases of lactic acid bacteria in Lactococcus lactis. Schwab C, Sørensen KI, and Gänzle MG Syst. Appl. Microbiol. 33(6), 300-7, (2010)

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Impact of manganese, copper and zinc ions on the transcriptome of the nosocomial pathogen Enterococcus faecalis V583. Abrantes, MC PLoS ONE 6(10), e26519, (2011)

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Scanning assay of beta-galactosidase activity. Li W, Zhao X, Zou S, et al. Prikl. Biokhim. Mikrobiol. 48(6), 668-72, (2012)

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Beta-galactosidase assay. Smale ST Cold Spring Harb. Protoc. 2010(5), pdb.prot5423, (2010)

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Methods for assessing the structure and function of cationic antimicrobial peptides. Pate M and Blazyk J Methods Mol. Med. 142, 155-73, (2008)

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Ceramic microsystem incorporating a microreactor with immobilized biocatalyst for enzymatic spectrophotometric assays. Baeza M, López C, Alonso J, et al. Anal. Chem. 82(3), 1006-11, (2010)

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A feeding tube model for activation of a cell-specific transcription factor during sporulation in Bacillus subtilis. Camp AH Genes Dev. 23(8), 1014-24, (2009)

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Overexpression and characterization of a novel transgalactosylic and hydrolytic β-galactosidase from a human isolate Bifidobacterium breve B24. Yi SH, Alli I, Park KH, et al. New Biotechnology 28(6), 806-13, (2011)

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Phage shock proteins B and C prevent lethal cytoplasmic membrane permeability in Yersinia enterocolitica. Horstman NK and Darwin AJ Mol. Microbiol. 85(3), 445-60, (2012)

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Characterization of the bga1-encoded glycoside hydrolase family 35 beta-galactosidase of Hypocrea jecorina with galacto-beta-D-galactanase activity. Gamauf C, Marchetti M, Kallio J, et al. FEBS J. 274(7), 1691-700, (2007)

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A novel metagenome-derived beta-galactosidase: gene cloning, overexpression, purification and characterization. Wang K, Li G, Yu SQ, et al. Appl. Microbiol. Biotechnol. 88(1), 155-65, (2010)

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Beta-galactosidase from Lactobacillus pentosus: purification, characterization and formation of galacto-oligosaccharides. Maischberger T, Leitner E, Nitisinprasert S, et al. Biotechnol. J. 5(8), 838-47, (2010)

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Functional interaction between herpes simplex virus type 2 gD and HVEM transiently dampens local chemokine production after murine mucosal infection. Yoon M, Kopp SJ, Taylor JM, et al. PLoS ONE 6(1), e16122, (2011)

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Role of the global transcriptional regulator PrrA in Rhodobacter sphaeroides 2.4.1: combined transcriptome and proteome analysis. Eraso JM J. Bacteriol. 190(14), 4831-48, (2008)

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Ser-796 of β-galactosidase (Escherichia coli) plays a key role in maintaining a balance between the opened and closed conformations of the catalytically important active site loop. Jancewicz LJ, Wheatley RW, Sutendra G, et al. Arch. Biochem. Biophys. 517(2), 111-22, (2012)

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Adverse effect of tannery waste leachates in transgenic Drosophila melanogaster: role of ROS in modulation of Hsp70, oxidative stress and apoptosis. Siddique HR, Gupta SC, Mitra K, et al. J. Appl. Toxicol. 28(6), 734-48, (2008)

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Interaction of Sesbania mosaic virus movement protein with the coat protein--implications for viral spread. Chowdhury SR and Savithri HS FEBS J. 278(2), 257-72, (2011)

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Differential expression of ccrA in methicillin-resistant Staphylococcus aureus strains carrying staphylococcal cassette chromosome mec type II and IVa elements. Higgins PG, Rosato AE, Seifert H, et al. Antimicrob. Agents Chemother. 53(10), 4556-8, (2009)

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A robust lentiviral pseudotype neutralisation assay for in-field serosurveillance of rabies and lyssaviruses in Africa. Wright E Vaccine 27(51), 7178-86, (2009)

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Purification and characterization of two phospho-β-galactosidases, LacG1 and LacG2, from Lactobacillus gasseri ATCC33323(T). Honda H, Nagaoka S, Kawai Y, et al. J. Gen. Appl. Microbiol. 58(1), 11-7, (2012)

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Characterization of lactose utilization and β-galactosidase in Lactobacillus brevis KB290, the hetero-fermentative lactic acid bacterium. Honda H, Yajima N, and Saito T Curr. Microbiol. 65(6), 679-85, (2012)

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The synergistic effect of Escherichia coli inactivation by sequential disinfection with low level chlorine dioxide followed by free chlorine. Yang W, Yang D, Zhu SY, et al. J. Water Health 10(4), 557-64, (2012)

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Activity modulation and reusability of beta-D-galactosidase confined in sol-gel derived porous silicate glass. Crescimbeni MC, Nolan V, Clop PD, et al. Colloids Surf. B Biointerfaces 76(2), 387-96, (2010)

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Production of 2-aminophenoxazin-3-one by Staphylococcus aureus causes false-positive results in β-galactosidase assays. Tse H, Chan E, Lam CW, et al. J. Clin. Microbiol. 50(11), 3780-2, (2012)

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Wide range of interacting partners of pea Gβ subunit of G-proteins suggests its multiple functions in cell signalling. Bhardwaj D, Lakhanpaul S, and Tuteja N Plant Physiol. Biochem. 58, 1-5, (2012)

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Highly regioselective galactosylation of floxuridine catalyzed by beta-galactosidase from bovine liver. Zeng QM, Li N, and Zong MH Biotechnol. Lett. 32(9), 1251-4, (2010)

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A new beta-galactosidase with a low temperature optimum isolated from the Antarctic Arthrobacter sp. 20B: gene cloning, purification and characterization. Białkowska AM, Cieśliński H, Nowakowska KM, et al. Arch. Microbiol. 191(11), 825-35, (2009)

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Practical considerations when using temperature to obtain rate constants and activation thermodynamics of enzymes with two catalytic steps: native and N460T-beta-galactosidase (E. coli) as examples. Kappelhoff JC, Liu SY, Dugdale ML, et al. Protein J. 28(2), 96-103, (2009)

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Investigation of factors affecting controlled release from photosensitive DMPC and DSPC liposomes. Aygun A, Torrey K, Kumar A, et al. Appl. Biochem. Biotechnol. 167(4), 743-57, (2012)

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Beil. 17,V,7,52

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