| Phosphate Assay 【Hannun/Obeid Lab 】 | 点击： 作者：51protocol收集 来源： 时间： 2007-03-18 本站论坛
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|  | Contributor: Jiandi Zhang
Date: July 4, 1996
1. Make standards using sodium phosphate at the following uM concentrations: 0, 2, 5, 7, 10, 20, 40, 60, and 80. Use the screw top glass tubes.
2. Dry the samples and standards under nitrogen evaporator (Rm 209), and add 150 ul 70% perchloric acid. Oxidize the samples and standards in a heating block (180C, Rm 218) overnight or for 1 hour if the heating block has been pre-heateed.
3. Rinse the sides of the tubes with 830ul xenopure water and vortex.
4. Add 170 ul 2.5% ammonium molybdate (by weight) and vortex.
5. Add 170 ul 10% ascorbic acid (by weight) and vortex. Make the ascorbic acid fresh every time.
6. Incubate for 15 min at 50°C. Allow tubes to cool.
7. Use spectrophotometer to read absorbance at 820nm.
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