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La cuantificación de la histidina quinasa bacteriana autofosforilación mediante un ensayo de nitrocelulosa Encuadernación
JoVE Revista
Bioquímica
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JoVE Revista Bioquímica
Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay
DOI:

09:17 min

January 11, 2017

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Capítulos

  • 00:05Título
  • 00:53Setting up the 96-well Dot Blot Apparatus
  • 02:02Reaction Initiation and Quenching
  • 02:57Spotting of Quenched Reactions on Nitrocellulose
  • 04:51Nitrocellulose Processing
  • 05:48Preparing Nitrocellulose Membrane for Scintillation Counting
  • 07:31Results: Quantification of the Autophosphorylation of V. parahaemolyticus H-NOX Associated Histidine Kinase (Vp1876)
  • 08:35Conclusion

Summary

Traducción Automática

We report and demonstrate an optimized nitrocellulose binding assay that can be used to quantify autophosphorylation of purified bacterial histidine kinases. Our method has several advantages over traditional SDS-PAGE based techniques, providing a valuable alternative for characterizing these important proteins.

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