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Cellular Membrane Affinity Chromatography Columns to Identify Specialized Plant Metabolites Interacting with Immobilized Tropomyosin Kinase Receptor B
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Cellular Membrane Affinity Chromatography Columns to Identify Specialized Plant Metabolites Interacting with Immobilized Tropomyosin Kinase Receptor B

Cellular Membrane Affinity Chromatography Columns to Identify Specialized Plant Metabolites Interacting with Immobilized Tropomyosin Kinase Receptor B

DOI:
10.3791/63118-v

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11:44 min

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January 19, 2022

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, , , ,
1Department of Biological Sciences,The University of Alabama, 2Department of Pharmaceutical Botany, Faculty of Pharmacy,Hacettepe University

Chapitres

  • 00:05Introduction
  • 00:52Cell Homogenization
  • 03:56Cell Membrane Solubilization
  • 04:46Cell Membrane Immobilization on IAM.PC.DD2 Particles
  • 06:23CMAC Column Packing
  • 08:57Results: Antibody Staining and Frontal Affinity Chromatography of Immobilized TrkB Receptors
  • 11:02Conclusion

Summary

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The protocol describes the preparation of cell membrane affinity chromatography (CMAC) columns with immobilized cell membrane fragments containing functional transmembrane tropomyosin kinase receptor B proteins. The use of CMAC columns in the identification of specialized plant metabolites interacting with these receptors and present in complex natural mixtures is also explained.

Tags

Cellular Membrane Affinity ChromatographyTransmembrane ReceptorsPlant MetabolitesTropomyosin Kinase ReceptorBenzamidinePMSFProtease InhibitorATPTris-HCl BufferHomogenization BufferCell HarvestingCell LysisColumn Preparation

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