ONPG(Synonyms: 2-Nitrophenyl β-D-galactopyranoside) | Cytiva思拓凡-Whatman滤纸滤膜滤器

生化分析试剂 Biochemical Assay Reagents
ONPG;(Synonyms: 2-Nitrophenyl β-D-galactopyranoside) 纯度: 99.84%

ONPG 是一种比色和分光光度计底物，用于检测 β-半乳糖苷酶活性。

ONPG Chemical Structure

CAS No. : 369-07-3

规格	价格	是否有货	数量
Free Sample (0.1-0.5 mg)	;	Apply now	;
10;mM;*;1 mL in Water	￥500	In-stock
500 mg	￥350	In-stock
1 g	￥500	In-stock
5 g	￥1000	In-stock
10 g	;	询价	;
50 g	;	询价	;

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ONPG 相关产品

bull;相关化合物库:

Bioactive Compound Library Plus
Glycoside Compound Library

生物活性

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

体外研究
(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 (K_m), turnover numbers (k_cat), and catalytic efficiencies (k_cat/K_m) of Gal308 for ONPG and lactose are determined. The k_cat and K_m 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 k_cat/K_m 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].

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

分子量

301.25

Formula

C₁₂H₁₅NO₈

CAS 号

369-07-3

运输条件

Room temperature in continental US; may vary elsewhere.

储存方式

Powder	-20deg;C	3 years
	4deg;C	2 years
In solvent	-80deg;C	6 months
	-20deg;C	1 month

溶解性数据

In Vitro:;

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

配制储备液

浓度溶剂体积质量	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
10 mM	0.3320 mL	1.6598 mL	3.3195 mL

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

参考文献

[1]. Zhang X, et al. Metagenomic approach for the isolation of a thermostable β-galactosidase with high tolerance of galactose and glucose from soil samples of Turpan Basin. BMC Microbiol. 2013 Oct 24;13:237. doi: 10.1186/1471-2180-13-237.

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.
参考文献	[1]. Zhang X, et al. Metagenomic approach for the isolation of a thermostable β-galactosidase with high tolerance of galactose and glucose from soil samples of Turpan Basin. BMC Microbiol. 2013 Oct 24;13:237. doi: 10.1186/1471-2180-13-237.

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.

参考文献

[1]. Zhang X, et al. Metagenomic approach for the isolation of a thermostable β-galactosidase with high tolerance of galactose and glucose from soil samples of Turpan Basin. BMC Microbiol. 2013 Oct 24;13:237. doi: 10.1186/1471-2180-13-237.