/
/
Forberedelse af grønkorn sub rum fra Arabidopsis til analyse af Protein lokalisering af Immunoblotting eller Proteomics
This content is Free Access.
Chapters
- 00:04Title
- 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
Her, vi beskriver en metode til at rense intakt grønkorn fra Arabidopsis blade og deres tre vigtigste sub rum (kuvert, stroma og tylakoiderne), ved hjælp af en kombination af differentieret centrifugations, kontinuerlig Percoll forløb, og diskontinuerlig saccharose forløb. Metoden er værdifuld for subplastidial og subcellulært lokalisering af proteiner af immunoblotting og proteomics.
