Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay

Jonathan Fischer; Roger A. Johnson; Elizabeth Boon

doi:10.3791/55129

Rivista

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Biochimica

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Quantificazione dei batteri istidina chinasi autophosphorylation usando un test nitrocellulosa Binding

Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay

JoVE Journal
Biochimica

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JoVE Journal Biochimica

Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay

Quantificazione dei batteri istidina chinasi autophosphorylation usando un test nitrocellulosa Binding

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8,484 Views

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09:17 min

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January 11, 2017

DOI:

10.3791/55129-v

DOI:

10.3791/55129-v

•

09:17 min

January 11, 2017

•

1 Views

Jonathan Fischer¹, Roger A. Johnson², Elizabeth Boon¹

¹Department of Chemistry,Stony Brook University, ²Department of Physiology and Biophysics,Stony Brook University

Trascrizione

Summary

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.