AMPK Methods and Protocols
concentration series of compounds. Total volume for each well is 200μl. The DMSO standards and the compound samples are set up f ...
The AMPK expression plasmid includes open reading frames of full-length α,β, and γsubunits of human AMPK with a ribosome-bindin ...
irreproducible results. Buffer pH should be adjusted and ver- ified using standard commercial pH meters and also double- checked ...
Determine the viral titer of the amplified viruses for each of the AMPK subunits. Any buffer that is free from high concentrati ...
multiple freeze/thaw cycles from the same stock to maintain protein stability. Solubility of AMPK ligands at high concentration ...
the observed conditions. While keeping the buffer (trisodium citrate at 100 mM) and ethylene glycol (1% (v/v)) concentra- tions ...
concentrations to get an initial estimate, and then repeat the experiment with at least eight concentrations ranging from 0.1 to ...
References Carling D (2017) AMPK signaling in health and disease. Curr Opin Cell Biol 45:31–37 Hardie DG, Schaffer BE, Brunet A ...
Li X, Wang L, Zhou XE, Ke J, de Waal PW, Gu X, Tan MH, Wang D, Wu D, Xu HE, Melcher K (2015) Structural basis of AMPK regulatio ...
Chapter 4 Biochemical Measurement of Glycogen: Method to Investigate the AMPK-Glycogen Relationship Elite Possik and Arnim Pause ...
residues are linked linearly with each other viaα(1-4) glycosidic bonds, and branching chains form every 8–12 residues viaα(1-6) ...
glycogen in the skeletal and cardiac muscles of pigs and mice [16–22]. Hunter et al. [14] demonstrated that the chronic AMPK act ...
2.2 Hydrolysis and Development Reagent Preparation In this section we provide the components that are utilized by most commercia ...
After boiling, take a 100μl aliquot for protein measurement (seeNote 4). Transfer the rest to a prechilled 15 ml conical tubes ...
For every sample include a sample background control. Add the same amount of samples used instep 3and complete the volume up to ...
foam. Cover plate with aluminum foil to protect from light, and incubate 30 min at room temperature with gentle shaking (seeTabl ...
ready-made kits where conditions are already optimized are important. However, based on the content of this chapter, where we de ...
and let it dry. Resuspend with Milli-Q H 2 O. Dilute the samples to be within the detection range. Dilution of the samples to m ...
Be careful while adding the hydrolysis enzyme mix. Make sure to add the 2μl inside the solution and not on the side and mix wel ...
Hunter RW, Treebak JT, Wojtaszewski JF, Sakamoto K (2011) Molecular mechanism by which AMP-activated protein kinase activation ...
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