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Chapters
- 00:05Title
- 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
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.
