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  4. ONPG

ONPG  (Synonyms: 2-Nitrophenyl β-D-galactopyranoside)

Cat. No.: HY-15926 Pureté: 99.91%
COA
SDS
Instruction de manipulation Technical Support

ONPG is a colorimetric and spectrophotometric substrate for detection of β-galactosidase activity.

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ONPG

ONPG Chemical Structure

CAS No. : 369-07-3

Size Prix Stock Quantité
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in Water
ready for reconstitution
 En stock
Solution
10 mM * 1 mL in Water En stock
Solid
500 mg En stock
1 g En stock
5 g En stock
10 g   Obtenir un devis  
50 g   Obtenir un devis  

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Based on 6 publication(s) in Google Scholar

Other Forms of ONPG:

ONPG Related Antibodies

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Description

ONPG is a colorimetric and spectrophotometric substrate for detection of β-galactosidase activity.

In Vitro

The enzyme displays high hydrolysis ability for ONPG (100%) and moderate activity for its natural substrate lactose (25.7%). However, the hydrolysis ability of the enzyme towards all other chromogenic nitrophenyl analogues is very weak, indicating that Gal308 is a β-galactosidase with narrow substrate specificity. To investigate the kinetic parameters of recombinant enzyme, the Michaelis-Menten constants (Km), turnover numbers (kcat), and catalytic efficiencies (kcat/Km) of Gal308 for ONPG and lactose are determined. The kcat and Km values are 464.7±7.8 s-1 and 2.7±0.3 mM for ONPG, and 264.2±2.1 s-1 and 7.1±0.8 mM for lactose, respectively. The kcat/Km value of the enzyme for ONPG (172.1 s-1mM-1) is 4.6-fold higher than that for lactose (37.2 s-1mM-1), which clearly demonstrated that the catalytic efficiency of Gal308 for ONPG is much higher than that for lactose[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

ONPG Related Antibodies
Masse moléculaire

301.25

Formule

C12H15NO8
CAS No.
Appearance

Solid

Color

White to off-white

SMILES

O[C@H]([C@@H](O)[C@H]1O)[C@@H](O[C@@H]1CO)OC2=C([N+]([O-])=O)C=CC=C2
Livraison

Room temperature in continental US; may vary elsewhere.
Stockage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 2 years
-20°C 1 year
Solvant et solubilité
In Vitro: 

H2O : 8.33 mg/mL (27.65 mM; ultrasonic and warming and heat to 60°C)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 3.3195 mL 16.5975 mL 33.1950 mL
5 mM 0.6639 mL 3.3195 mL 6.6390 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

  • Calculateur de molarité

  • Calculateur de dilution

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo Dissolution Calculator
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Working solution concentration: mg/mL
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only.If necessary, please contact MedChemExpress (MCE).
Pureté et documentation

Purity: 99.91%

SDS (393 KB)

COA (255 KB) HNMR (268 KB) LCMS (93 KB)

Instruction de manipulation (2659 KB)
Références
Essai de kinase
[1]

The β-galactosidase activity is measured using two substrates including ONPG and lactose in this study. The β-galactosidase activity for ONPG is measured by following the amount o-nitrophenol released from ONPG. The reaction mixture is composed of 100 μL of the enzyme solution and 400 μL of ONPG solution (2.5 g/L in 100 mM Tris-HCl buffer at pH 6.8). After incubation at 78°C for 15 min, the reaction is terminated by adding an equal volume of 1 M Na2CO3. The released o-nitrophenol is quantitatively determined by measuring at A405. One unit of activity is defined as the amount of enzyme needed to produce 1 μmol of o-nitrophenol per minute under the assay condition. The specific activity is expressed as units per milligram of protein. Assays for activity towards lactose are performed in the same buffer containing 100 μL of enzyme solution and 5% lactose, and the reaction is stopped by boiling for 10 min, and the concentration of glucose is determined using a glucose oxidase-peroxidase assay kit. The released glucose is quantitatively determined by measuring A492. One unit of enzyme activity is defined as the amount of activity required to release 1 μmol of glucose per minute[1].

MCE n'a pas confirmé de manière indépendante l'exactitude de ces méthodes. Ils sont pour référence seulement.
Références

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
H2O 1 mM 3.3195 mL 16.5975 mL 33.1950 mL 82.9876 mL
5 mM 0.6639 mL 3.3195 mL 6.6390 mL 16.5975 mL
10 mM 0.3320 mL 1.6598 mL 3.3195 mL 8.2988 mL
15 mM 0.2213 mL 1.1065 mL 2.2130 mL 5.5325 mL
20 mM 0.1660 mL 0.8299 mL 1.6598 mL 4.1494 mL
25 mM 0.1328 mL 0.6639 mL 1.3278 mL 3.3195 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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ONPG Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

ONPG369-07-32-Nitrophenyl β-D-galactopyranosideBiochemical Assay ReagentsInhibitorinhibitorinhibit

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