The depurination activity of mucoricin was measured by the release of adenine when mammalian RNA was treated with mucoricin for 24 h at 37oC32 (link). Mammalian RNA extracted from A549 alveolar epithelial cells by Qiagen RNasy mini kits according to manufacturer’s instruction were treated with 20 μg/ml of mucoricin in 0.01M HEPES/10 mM ammonium acetate buffer containing 1 mg/ml BSA for 24 h at 37oC. The solution was then filtered through a 10 kDa size exclusion column and 40 μl was injected into HPLC using Phenomenex Luna C18 reverse phase column (10 × 250 mm) attached to a Varian ProStar HPLC 218 system (Varian, Walnut Creek, CA). Solvent A was 20 mM ammonium acetate, and solvent B was 100% acetonitrile. The column gradient was as follows: 97% to 60% solvent A in 10 min at flow rate of 1 ml/min. The column effluent was monitored at 260 nm.
Prostar hplc 218 system
Manufactured by Agilent Technologies
The ProStar HPLC 218 system is a high-performance liquid chromatography (HPLC) instrument designed for analytical and preparative applications. It features a modular design, allowing for customization to meet specific research or laboratory requirements. The system includes a solvent delivery module, an autosampler, a UV-Vis detector, and a data management system.
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Lab products found in correlation
2 protocols using prostar hplc 218 system
1
Quantifying Mucoricin-Mediated RNA Depurination
2
Quantifying Mucoricin-Mediated RNA Depurination
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The depurination activity of mucoricin was measured by the release of adenine when mammalian RNA was treated with mucoricin for 24 h at 37oC32 (link). Mammalian RNA extracted from A549 alveolar epithelial cells by Qiagen RNasy mini kits according to manufacturer’s instruction were treated with 20 μg/ml of mucoricin in 0.01M HEPES/10 mM ammonium acetate buffer containing 1 mg/ml BSA for 24 h at 37oC. The solution was then filtered through a 10 kDa size exclusion column and 40 μl was injected into HPLC using Phenomenex Luna C18 reverse phase column (10 × 250 mm) attached to a Varian ProStar HPLC 218 system (Varian, Walnut Creek, CA). Solvent A was 20 mM ammonium acetate, and solvent B was 100% acetonitrile. The column gradient was as follows: 97% to 60% solvent A in 10 min at flow rate of 1 ml/min. The column effluent was monitored at 260 nm.
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