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Identifying the Binding Proteins of Small Ligands with the Differential Radial Capillary Action of Ligand Assay (DRaCALA)
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Journal JoVE Biochimie
Identifying the Binding Proteins of Small Ligands with the Differential Radial Capillary Action of Ligand Assay (DRaCALA)

Identifying the Binding Proteins of Small Ligands with the Differential Radial Capillary Action of Ligand Assay (DRaCALA)

DOI:
10.3791/62331-v

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09:26 min

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March 19, 2021

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, ,
1Department of Biology,University of Copenhagen

Chapitres

  • 00:05Introduction
  • 00:42Preparation of Whole Cell Lysates
  • 01:43Nickel-Nitrilotriacetic Acid (Ni-NTA) Affinity Purification of Relseq and GppA
  • 02:55Gel Filtration
  • 03:33Synthesis of 32P-Labeled Guanosine Pentaphosphate (pppGpp) and Guanosine Tetraphosphate (ppGpp)
  • 05:08DRaCALA Screening of the Target Proteins of (p)ppGpp
  • 06:37Quantification and Identification of Potential Target Proteins
  • 07:46Results: DRaCALA Plate Screening and Quantification
  • 08:39Conclusion

Summary

Traduction automatique

Traduction automatique

The Differential Radial Capillary Action of Ligand Assay (DRaCALA) can be used to identify small ligand binding proteins of an organism by using an ORFeome library.

Tags

Keywords: DRaCALASmall Signaling MoleculesEffector ProteinsOVium LibraryE. ColiIPTGLysis BufferNickel-NTA ResinWashing BufferElution BufferGel Filtration

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