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拟南芥叶绿体子隔层的制备及免疫印迹蛋白质组化分析
Journal JoVE
Biochimie
This content is Free Access.
Journal JoVE Biochimie
Preparation of Chloroplast Sub-compartments from Arabidopsis for the Analysis of Protein Localization by Immunoblotting or Proteomics
DOI:

10:28 min

October 19, 2018

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Chapitres

  • 00:04Titre
  • 01:01Harvesting of Arabidopsis Leaves
  • 02:26Purification of Crude Chloroplasts Using Differential Centrifugation
  • 03:30Purification of Intact Chloroplasts on Continuous Percoll Gradient
  • 04:51Lysis of Intact Chloroplasts Using a Hypotonic Buffer and Purification of Chloroplast Sub-Compartments on Discontinuous Sucrose Gradients
  • 06:05Washing and Concentration of Thylakoid and Envelope Membrane Systems
  • 07:09Results: Analysis of Protein Localization by Immunoblotting or Proteomics
  • 08:44Conclusion

Summary

Traduction automatique

在这里, 我们描述了从拟南芥叶及其三个主要子隔间 (包络、基质和 thylakoids) 中纯化完整叶绿体的方法, 使用微分 centrifugations、连续 Percoll 梯度的组合和不连续的蔗糖梯度。该方法对蛋白质的 subplastidial 和亚细胞定位具有重要的免疫印迹和蛋白质组学意义。

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