AMPK Methods and Protocols

7 mL Scintillation vials (or equivalent). Scintillation counter (e.g., Tri-Carb 2910 TR, Perkin Elmer). Tabletop centrifuge cap ...

Spin down the media at 1500gfor 10 min to remove any detached cells. Pool the medium collected from all flasks and filter-ster ...

Plate bone marrow cells in 100 mm tissue culture plates (10 mL per plate) (seeNote 8). Incubate the cells in a humidified 5% CO ...

Visualize the TLC plate by transferring plate to a new TLC tank containing iodine pellets (or by preferred visualization techni ...

Enter the average absorbance value from each experimental duplicate into the rearranged equation (as they-value), and then proc ...

Add 1μL of 50 mM A-769662 stock for a final concentration of 100μM or desired AMPK activator. Incubate in a humidified 5% CO 2 ...

Incubate cells in a humidified 5% CO 2 incubator at 37C for 30 h. Cells can be treated simultaneously with AMPK activators (se ...

4 Notes Large volumes of L929-conditioned medium can be obtained by simply scaling up the number of T175 flasks. Typical batche ...

Longer time points could potentially skew interpretation given the metabolism of lipids into which^3 H-acetate has been incorpo ...

In vivo reverse cholesterol transport assay can employ the use of wild-type or AMPK-deficient cells. In addition, AMPK acti- va ...

Larach DB, deGoma EM, Rader DJ (2012) Targeting high density lipoproteins in the pre- vention of cardiovascular disease? Curr C ...

Chapter 31 Modulation of Vascular Function by AMPK: Assessment of NO Bioavailability and Surrogates of Oxidative Stress Swenja K ...

in the case oflNO) is used for spin trapping as this more stable adduct formation allows measurement oflNO at a specific point i ...

analytical grade reagents. Prepare and store all reagents at room temperature (unless indicated otherwise). Diligently follow al ...

an autosampler (AS-2057 plus) from Jasco (Groß-Umstadt, Germany), and a C 18 Nucleosil 100-3 (125/4) column from Macherey and Na ...

3 Methods Carry out all procedures at room temperature unless otherwise specified. 3.1 Vascular Nitric Oxide Analysis by EPR Spe ...

Transfer the tube to the EPR machine and start the measure- ment. Figure1 shows representative EPR tracings from murine aortic ...

After 20 min remove the samples from the heater at the same time, and transfer the solution to the HPLC vials. Make sure that t ...

3.3.1 Sample Preparation Remove all connective tissue as well as vascular fat tissue from aortic sections. Dissect the aorta in ...

3.4.2 Preparation of Tissue Samples Homogenate frozen tissue samples on liquid nitrogen using a mortar and pestle. Tissue shoul ...