Journal
/
生物化学
/
Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay
JoVE 杂志
生物化学
需要订阅 JoVE 才能查看此.  登录或开始免费试用。
JoVE 杂志 生物化学
Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay

Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay

8,408 Views

•

09:17 min

•

January 11, 2017

DOI:

10.3791/55129-v

DOI:
10.3791/55129-v

•

09:17 min
January 11, 2017

•

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

Summary

Automatically generated

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.

Explore More Videos

Bacterial Histidine KinaseAutophosphorylationQuantificationNitrocellulose Binding AssayTwo-component SignalingPost-translational ModificationRadioactive LigandsHigh-throughputKinetic ParametersProtein-protein InteractionsFiltration96-well Dot-plot ApparatusGamma-32P ATPPhosphoric Acid

Read Article

Cite this Article

Fischer, J., Johnson, R. A., Boon, E. Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay. J. Vis. Exp. (119), e55129, doi:10.3791/55129 (2017).

Download .ris file

Copy

Share Video

.

Copy