Membrane templates were first equilibrated in filtered and degassed assay buffer, after which, 200 μl of assay buffer containing 1% bovine serum albumin (Sigma-Aldrich, St. Louis, MO) was flowed in and incubated for 10 min. Subsequently, 200 μl of 200 nM each adaptor (diluted in assay buffer) was introduced and incubated with templates for 10 min. Excess adaptor was rinsed with 2 × 200 μl of assay buffer and 1 × 200 μl of assay buffer containing an oxygen scavenger cocktail of 0.2 mg/ml glucose oxidase (G-2133; Sigma-Aldrich), 0.035 mg/ml catalase (C-40; Sigma-Aldrich), 4.5 mg/ml glucose, and 1 mM dithiothreitol. The clathrin assembly reaction was monitored in real time by flowing in 200 μl of 80 nM Texas red–labeled clathrin and freshly diluted in assay buffer containing the oxygen scavenger cocktail. All reactions were carried out at 25°C.
Glucose oxidase g 2133
Manufactured by Merck Group
Glucose oxidase (G-2133) is a lab equipment product provided by Merck Group. It is an enzyme that catalyzes the oxidation of glucose to gluconic acid and hydrogen peroxide. The core function of this product is to serve as a reagent for the quantitative determination of glucose levels.
Automatically generated - may contain errors
2 protocols using glucose oxidase g 2133
1
Real-Time Monitoring of Clathrin Assembly
2
Preparation of Mock Sputum and Bacterial Cultures
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Orange-red food dye was obtained from Ajanta Industries (Gurugram, India); papers were dipped in a solution of 2 g/L and dried in an incubator at 37 °C; for preparation of mock sputum: methylcellulose (M-0262) and egg yolk emulsion (17148) were obtained from Sigma-Aldrich; mock sputum was prepared as a solution of 1.8% w/v methylcellulose and 10% egg yolk emulsion in DI water21 (link); for Mycobacterial culture: Tryptone soya broth 24392 (TM 018) was obtained from SRL and Middlebrook 7H9 Broth Base (M198-500G) was obtained from HiMedia. Tween-80 28940 (1628157) and glucose 51758 (0449130), obtained from SRL, were used as supplements; for E.coli culture: Luria Bertani broth, Lennox 14593 (LM 021) and agar powder 85473 (014042) were obtained from SRL; for staining bacterial colonies: ethidium bromide (EtBr) was obtained from SRL; for glucose detection assay: dextrose (AR 51758) was obtained from SRL and the following reagents were obtained from Sigma: glucose oxidase (G2133), horseradish peroxidase (P8375), 4-amino antipyrine (A4382) and 2,4,6-tribromo-3-hydroxybenzoic acid (439533).
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