For purification of the SalC assay product, reactions were set up as described above at a 50 mL scale in an Erlenmeyer flask. SalB-PCP loading was allowed to proceed for 12 hours, at which point approximately 1 gram of XAD7 resin and SalC (20 μM) were added to the reaction mixture and assays were allowed to proceed overnight. In the morning, resin was extracted (3×) with ethyl acetate. Organic extracts were combined and evaporated to dryness. Samples were resuspended in acetonitrile and the peak corresponding to R-17 was purified by preparative HPLC using a Phenomenex Luna C18 column (5 μm, 100 mm, 2mm i.d.), along with an Agilent Technologies system composed of a PrepStar pump, a ProStar 410 autosampler, and a ProStar UV detect (Agilent Technologies, Inc, Santa Clara, USA). The sample were eluted by a gradient from 20–100% acetonitrile over 40 min at a flow rate of 10 mL/min. The peak corresponding to R-17 was collected and dried by rotary evaporation and lyophilization. Finally, R-17 was purified using a small-scale silica column with a dichloromethane/acetonitrile solvent system. All stages of purification were monitored by LCMS analysis.
Prostar uv detect
Manufactured by Agilent Technologies
Sourced in United States
The ProStar UV detect is a laboratory instrument designed for the detection and analysis of compounds using ultraviolet (UV) light. It serves as a key component in various analytical procedures, providing accurate and reliable UV detection capabilities.
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2 protocols using prostar uv detect
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Purification of SalC Assay Product
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Purification of SalC Assay Product
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For purification of the SalC assay product, reactions were set up as described above at a 50 mL scale in an Erlenmeyer flask. SalB-PCP loading was allowed to proceed for 12 hours, at which point approximately 1 gram of XAD7 resin and SalC (20 μM) were added to the reaction mixture and assays were allowed to proceed overnight. In the morning, resin was extracted (3×) with ethyl acetate. Organic extracts were combined and evaporated to dryness. Samples were resuspended in acetonitrile and the peak corresponding to R-17 was purified by preparative HPLC using a Phenomenex Luna C18 column (5 μm, 100 mm, 2mm i.d.), along with an Agilent Technologies system composed of a PrepStar pump, a ProStar 410 autosampler, and a ProStar UV detect (Agilent Technologies, Inc, Santa Clara, USA). The sample were eluted by a gradient from 20–100% acetonitrile over 40 min at a flow rate of 10 mL/min. The peak corresponding to R-17 was collected and dried by rotary evaporation and lyophilization. Finally, R-17 was purified using a small-scale silica column with a dichloromethane/acetonitrile solvent system. All stages of purification were monitored by LCMS analysis.
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