AMPK Methods and Protocols

4 Notes

1. After adjusting the pH, the solution turns to pale green color.


2. If stored at 4C, use the reagent within 15 days. You can make
   aliquots and store them at 20 C for longer preservation.


3. Store in ready to use aliquots. Preheat (95C) before use to
   solubilize the reagent.


4. It is important to habituate the animals to handling before ICV
   injections; thus, for acute procedures, the stress induced during
   the injection is minimized.


5. Due to the biological features of adenoviruses, the infection
   and overexpression are immediate, but the effect is transitory,

Fig. 2Western blot autoradiographic images. (a) Autoradiographic images of

pAMPKα, AMPKα1, AMPKα2, andβ-actin from hypothalamic samples of rats

separated by 6.5% SDS-PAGE electrophoresis. (b) Representative

autoradiographic images of pAMPKα, AMPKα1, AMPKα2, andβ-actin from

hypothalamic samples of fed and fasted rats separated by 6.5% SDS-PAGE

electrophoresis. Vertical lines show spliced bands coming in all the cases from

the same gels [61]

444 Patricia Seoane-Collazo and Miguel Lo ́pez