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The standardized procedure used with GL Sciences' Phos-TiO Kits consists of four steps with a total processing time typically under 40 minutes. First, the Phos-TiO spin columns are conditioning using the supplied buffer, the sample is then applied, and then rinsed using the supplied rinsing buffer. Purified phosphopeptides are then eluted using an MS-friendly 0.5% ammonium hydroxide solution. Each step of the process is performed using a centrifuge, and Phos-TiO kits require reusable adaptors to secure the spin columns in the waste and collection tubes. (As these adaptors are reusable, they are not components of each kit and much be ordered, once, separately from the kits). Adaptors are the same for individual 10uL and 200uL tubes, while separate 96-well plate adaptors are available for 10uL and 200uL tips, as shown here. 
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