Analysis of the Incorporation of Carbon Atoms from Radioactive Lactate into Proteins
碳原子从放射性乳酸盐掺入到蛋白质的分析
发布: 2013年07月05日第3卷第13期 DOI: 10.21769/BioProtoc.812 浏览次数: 7107
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参见作者原研究论文
The authors used this protocol in:
Oct 2012
Abstract
This method allows to analyze if the carbon atoms of lactate are embedded into proteins. Indeed, mammalian cells express the transporter of monocarboxylic acids (called MCT1) that allows the entry of lactate into the cell. To this end, cells are incubated for 24 h with the culture medium containing lactate uniformly labeled with carbon 14 and then, lactate inside the cell is evaluated by counting the radioactivity by a scintillator.
Materials and Reagents
- [U-14C] lactate (U: uniformly labelled) (PerkinElmer, catalog number: NEC599250UC )
- Phosphate buffered saline (pH 7) (Sigma-Aldrich, catalog number: D8537 )
- Trichloroacetic acid (Sigma-Aldrich, catalog number: T6399 )
- Liquid scintillation (PerkinElmer, catalog number: 6NE9529 )
- Vials for scintillator (PerkinElmer)
- Dulbecco’s Modified Eagle Medium (DMEM)
- Eppendorf tube
- Water
- Cell scrapers (Euroclone)
Equipment
- Incubator for cell culture
- Petri dish for cell culture (six inches diameter)
- Scintillator
- Burker chamber
Procedure
文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Fiaschi, T. and Chiarugi, P. (2013). Analysis of the Incorporation of Carbon Atoms from Radioactive Lactate into Proteins . Bio-protocol 3(13): e812. DOI: 10.21769/BioProtoc.812.
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分类
生物化学 > 糖类 > 乳酸
细胞生物学 > 细胞新陈代谢 > 糖类
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