The carcinogen 1, 2-dimethylhydrazine (DMH) was obtained from Tokyo Chemical Industry. P-nitrophenyl-β-d -glucuronide, E coli β-glucuronidase, glutathione, uridine diphosphate glucuronic acid (UDPGA), 1-chloro-2,4dinitro benzene (CDNB), glycogen, guanine and O6-MeG were purchased from Sigma-Aldrich Chemical Co. Tris–HCl and sucrose were obtained from Thermo Fisher Scientific Inc. Phenol–chloroform–isoamylalcohol was obtained from Research Organics Inc. Other reagent grade chemicals were obtained from Merck Millipore Bioscience (Thailand).
E coli β glucuronidase
Manufactured by Merck Group
Sourced in Thailand
E. coli β-glucuronidase is an enzyme derived from the bacterium Escherichia coli. It catalyzes the hydrolysis of β-D-glucuronides. This enzyme is commonly used in various analytical and research applications.
Automatically generated - may contain errors
Lab products found in correlation
O6 meg, by Merck Group (2 mentions)
1 chloro 2 4dinitro benzene cdnb glycogen, by Merck Group (2 mentions)
Uridine diphosphate glucuronic acid udpga, by Merck Group (2 mentions)
Glutathione (gsh), by Merck Group (2 mentions)
Guanine, by Merck Group (2 mentions)
P nitrophenyl β d glucuronide, by Merck Group (2 mentions)
Sucrose, by Thermo Fisher Scientific (1 mentions)
Tris hcl, by Thermo Fisher Scientific (1 mentions)
Bovine hemin, by Merck Group (1 mentions)
Bbl brain heart infusion medium, by BD (1 mentions)
Dulbecco s phosphate buffered saline, by Thermo Fisher Scientific (1 mentions)
Vitamin k1, by Merck Group (1 mentions)
Milli q purification system, by Merck Group (1 mentions)
Amoxapine, by Merck Group (1 mentions)
P nitrophenol, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Agar, by Biosharp (1 mentions)
4 protocols using e coli β glucuronidase
1
Carcinogen-Induced Enzymatic Assay
2
Enzymatic Prodrug Hydrolysis Analysis
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Hydrolysis of the prodrugs was carried out by incubating the prodrugs at a final concentration of (20 μg/mL) with e.coli β-glucuronidase (Sigma, G 7646) (1.0 μg/mL) in 10 mM PBS buffer solution pH = 7.4. Aliquots were taken out at given time points (30 min, 120 min, 1 d), enzyme was precipitated in cold MeOH (2 mL), and the samples were centrifuged (1400 rcf, 4 °C, 5 min). The supernatant was transferred, the solvent was removed in vacuo, and the sample dissolved in 50 μL H2O/MeCN (95/05), and analyzed via HPLC. Method A: Elution was performed starting with solvent B 5% to B 100% over 15 min at T = 40 °C at a flow rate of 0.4 mL/min. Detection was performed by UV detector (254 nm and 280 nm). Stability tests were carried out according to the hydrolysis protocol, without the addition of enzyme.
3
Enzymatic Assay for β-Glucuronidase
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DMSO, bovine hemin, vitamin K1, amoxapine, p-nitrophenol (PNP), p-nitrophenyl-β-D-glucuronide (PNPG), and E. coli β-glucuronidase (G8401) were supplied by Sigma-Aldrich (St. Louis, MO). L-cystine was purchased from Research Organics (Cleveland, OH). Purities were all >98%. Agar was purchased from Biosharp (Hirono, Japan). The BBL Brain Heart Infusion (BHI) medium was purchased from Becton Dickinson (Franklin Lakes, NJ). Dulbecco's phosphate-buffered saline (PBS) was provided by Life Technologies (Carlsbad, CA). Deionized water was purified by a Milli-Q purification system (Millipore, Bedford, MA).
4
Carcinogen DMH Protocol for Bioassays
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The carcinogen 1, 2-dimethylhydrazine (DMH) was obtained from Tokyo Chemical Industry. P-nitrophenylβ-D-glucuronide, E coli β-glucuronidase, glutathione, uridine diphosphate glucuronic acid (UDPGA), 1chloro-2,4dinitro benzene (CDNB), glycogen, guanine and O 6 -MeG were purchased from Sigma-Aldrich Chemical Co. Tris-HCl and sucrose were obtained from Thermo Fisher Scienti c Inc. Phenol-chloroformisoamylalcohol was obtained from Research Organics Inc. Other reagent grade chemicals were obtained from Merck Millipore Bioscience (Thailand).
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