Difference between revisions of "IC4R011-Proteomic-1999-10217180"
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==Protein Extraction and 2-D PAGE== | ==Protein Extraction and 2-D PAGE== | ||
| + | *Following separation by 2-D PAGE, the proteins were electroblotted onto a polyvinylidene difluoride (PVDF)membrane (ProBlott; Applied Biosystems, Foster City,CA, USA) and detected by Ponceau 3R staining [11]. The spots were excised from the PVDF membrane and applied to the upper glass block of the reaction chamber in a gas-phase protein sequencer (477A; Applied Biosystems). Edman degradation was performed according to the standard program supplied by Applied Biosystems.The released phenylthiohydantoin amino acid derivatives were identified by the on-line systems of high performance liquid chromatography (120A; Applied Biosystems). | ||
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==Research Findings== | ==Research Findings== | ||
Revision as of 14:17, 17 July 2016
Contents
Project Title
- Separation and characterization of proteins from green and etiolated shoots of rice (Oryza sativa L.):Towards a rice proteome
The Background of This Project
Plant Culture & Treatment
- Green and etiolated shoots of rice (Oryza sativa L. cultivar Nipponbare) were used for this study. Rice seeds were sown on vermiculite and incubated at 25oC under light for 12 h and in the dark for 12 h (for green shoots) or in the dark for 24 h (for etiolated shoots)
Protein Extraction and 2-D PAGE
- Following separation by 2-D PAGE, the proteins were electroblotted onto a polyvinylidene difluoride (PVDF)membrane (ProBlott; Applied Biosystems, Foster City,CA, USA) and detected by Ponceau 3R staining [11]. The spots were excised from the PVDF membrane and applied to the upper glass block of the reaction chamber in a gas-phase protein sequencer (477A; Applied Biosystems). Edman degradation was performed according to the standard program supplied by Applied Biosystems.The released phenylthiohydantoin amino acid derivatives were identified by the on-line systems of high performance liquid chromatography (120A; Applied Biosystems).
Research Findings
Labs working on this Project
- Department of Molecular Genetics, National Institute of Agrobiological Resources,Tsukuba, Japan
Corresponding Author
- Dr. Setsuko Komatsu:skomatsu@abr.affrc.go.jp
