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Phloem Sap Sampling from Brassica napus for 3D-PAGE of Protein and Ribonucleoprotein Complexes
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Biology
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Phloem Sap Sampling from Brassica napus for 3D-PAGE of Protein and Ribonucleoprotein Complexes

Phloem Sap Sampling from Brassica napus for 3D-PAGE of Protein and Ribonucleoprotein Complexes

DOI:
10.3791/57097-v

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11:23 min

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January 09, 2018

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, , ,
1Molecular Plant Genetics,Universität Hamburg

Chapters

  • 00:05Title
  • 01:04Sampling and Preparation of Phloem Sap from B. napus Plants
  • 02:27Blue Native PAGE
  • 03:37OPTIONAL: Detection of RNA Using Blue Native Northern Blotting
  • 05:30Second Dimension: Tris-tricine-urea PAGE
  • 07:29Third Dimension: SDS-PAGE
  • 08:40Results: Analysis of Protein Composition of B. napus Phloem Exudate
  • 10:06Conclusion

Summary

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Here we present a protocol to analyze the protein composition of large native protein:protein and protein:nucleic acid complexes from oilseed rape (B. napus) phloem exudate using a 3D polyacrylamide gel electrophoresis (PAGE) approach combining blue native (BN) with two denaturing PAGEs followed by mass spectrometric identification.

Tags

Keywords: Phloem Sap3D-PAGEBrassica NapusProteinRibonucleoprotein ComplexesBlue Native PAGEProtein-protein InteractionsProtein-nucleic Acid InteractionsOilseed RapeCentrifugal ConcentrationElectrophoresis

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