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  4. ONPG

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

Cat. No.: HY-15926 Purity: 99.84%
COA Handling Instructions

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

For research use only. We do not sell to patients.

ONPG Chemical Structure

ONPG Chemical Structure

CAS No. : 369-07-3

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Solid + Solvent (Highly Recommended)
10 mM * 1 mL in Water
ready for reconstitution
 USD 33 In-stock
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10 mM * 1 mL in Water USD 33 In-stock
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5 g USD 80 In-stock
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Based on 4 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
Molecular Weight

301.25

Formula

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
Shipping

Room temperature in continental US; may vary elsewhere.
Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 2 years
-20°C 1 year
Solvent & Solubility
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.

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  • Dilution Calculator

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

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

This equation is commonly abbreviated as: C1V1 = C2V2

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In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Calculation results:
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).
Purity & Documentation

Purity: 99.84%

COA (255 KB) HNMR (253 KB) LCMS (265 KB)

Handling Instructions (2659 KB)
References
Kinase Assay
[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 has not independently confirmed the accuracy of these methods. They are for reference only.
References

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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