AMPK Methods and Protocols
it is then straightforward to calculate Gibbs free energy (ΔG) and the entropy (ΔS). The following relationships are required to ...
these experiments are conducted on isotope (^15 Nor^13 C)-enriched molecules, and for that reason protein, rather than ligand, r ...
transferred into the z-axis (alongB 0 ) with varying mixing times (Tmix) at which point interconversion between states is allowe ...
1200 Hz) where theR 2 obsis dependent onνCPMG(Fig.3). The intensity of the peaks can be taken and converted to plots ofR 2 obsvs ...
The combination of ITC and NMR is a proven method and has been used previously for studying protein interactions when con- venti ...
3 Methods 3.1 Sample Preparation for ITC and NMR Spectroscopy Before ITC experiments, unlabelledβ1- andβ2-CBM protein samples a ...
The sample is gradient-shimmed, optimally using a round of three-dimensional shimming and then one-dimensional shim- ming until ...
As for the ZZ-exchange experiment, the sample is tuned and shimmed, and the^1 H90pulse is determined. A list of pulse frequenc ...
have been assigned. For any ligand binding study, it is desirable to assign at least the backbone (^1 HN,^15 N,^13 Cα,^13 Cβ) re ...
-xSW 10416.667 -zSW 2351.499 \ -xOBS 800.133 -zOBS 81.086 \ -xCAR 4.703 -zCAR 119.000 \ -xLAB HN -zLAB 15N \ -ndim 3 -aq2D State ...
Chapter 7 Bioinformatics Approach to Identify Novel AMPK Targets Brendan Gongol, Traci Marin, David A. Johnson, and John Y.-J. S ...
the kinase and substrate. Other pattern matching algorithms, including Scansite, utilize position weight matrices to create a pr ...
Purified recombinant target protein. Target protein can be purified from cell lysate or purchased commercially. Store at 80 ...
Autoradiography film: commercially available. Film fixative and developer: commercially available. Automated film developer (op ...
(K|R|H)(L|V|M|I|A|F)(K|R|H)...(S|T)...(I|L|V|M|A|F)", proteomehuman)] Remove any duplicated hits by running the following code ...
3.2 Validation of Putative AMPK Targets All procedures are to be performed at room temperature unless otherwise specified. Prep ...
second piece of transparency sheet over the gel and then secure with laboratory tape (Fig.2). (Optional) Use gel drying equipme ...
2 weeks depending on the radiant intensity of the sample (see Note 19). Following exposure, the films can be developed in a dar ...
The error “Error in library() : there is no package called “” indicates that the package is not installed. Rerun the code liste ...
SDS-PAGE gel percentage must be optimized to ensure max- imum separation between AMPK subunits and target proteins of interest. ...
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