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Extra resources for Carbon-Carbon Composite Molten-Salt Cooled Space Reactors [pres. slides]
7 by KOH. This medium formula was modified from MS medium and specially designed for Arabidopsis growing on solid medium in plate or jar. The medium should be made in two separate sets and labeled clearly as a light nitrogen medium (14N, from normal KNO3) or a heavy nitrogen medium (15N, from K15NO3). 2. Protein Sample Preparations 1. 2% TritonX-100, 20 mM EDTA, 20 mM EGTA, 50 mM NaF, 2 mM NaVO3, and 1% Glycerol 2-phosphate disodium salt hydrate, stored at 4 C. 5% phosphatase inhibitors cocktail 2 (and phosphatase inhibitors cocktail 1, which can be purchased only in some regions and countries), 1Â complete EDTA-free protease inhibitors cocktail, 5 mM ascorbic acid, and 2% PVPP.
Protein Sample Preparation 1. Seedlings (10 g) are ground into powders with liquid nitrogen in a precooled mortar. To effectively denature plant proteins for the purpose of freezing phosphor group onto phosphosite and to prevent in vitro nonspecific enzyme catalysis by kinase and phosphatase during protein preparation, a phosphoprotein extraction buffer UEB is employed during plant protein isolation. The tissue powder is then mixed with 50 ml UEB buffer and ground for 2 min. 4 Sorvall rotor). The high-speed centrifugation is designed to remove cell debris, DNA, cell wall, lipid, and RNA.
Nucleic Acids Res 34:3267–3278 7. Chitteti BR, Peng Z (2007) Proteome and phosphoproteome dynamic change during cell dedifferentiation in Arabidopsis. Proteomics 7:1473–1500 8. Li X, Gerber SA, Rudner AD, Beausoleil SA et al (2007) Large-scale phosphorylation analysis of a-factor-arrested Saccharomyces cerevisiae. J Proteome Res 6:1190–1197 9. Ono M, Shitashige M et al (2006) Label-free quantitative proteomics using large peptide data sets generated by nanoflow liquid chromatography and mass spectrometry.
Carbon-Carbon Composite Molten-Salt Cooled Space Reactors [pres. slides]