AMPK Methods and Protocols

3.4.4 Dot Blot 1. Prepare the filter paper and the nitrocellulose membrane for dot botting by equalizing both in PBS for 5 min. ...

Expose to film or analyze with a chemiluminescent detection system. For quantification purposes, a densitometric analysis of ea ...

Zou M, Martin C, Ullrich V (1997) Tyrosine nitration as a mechanism of selective inactivation of prostacyclin synthase by perox ...

Chapter 32 Measurement of Reactive Oxygen Species (ROS) and Mitochondrial ROS in AMPK Knockout Mice Blood Vessels Qilong Wang an ...

progression of CVD, including atherosclerosis, hypertension, and heart failure [1]. Therefore, alterations to physiologic ROS le ...

HPLC-fluorescence protocol that allows specific measurement of mitochondrial O 2 l
using MitoSOX. 2-OH-mito-E+is identified and ...

In this chapter, we describe a novel fluorescence dye-based O 2 l
and mitochondrial O 2 l
measurement method in blood ves- sels ...

MitoSOX staining solution: 2μM MitoSOX dissolved in Krebs buffer (seeNote 1). 50% methanol: Methanol diluted in Milli-Q pure H ...

Remove the heart and fat tissue around the aortic region without cutting aortic wall under dissecting microscope (see Note 3). ...

3.3 Measurement of Mitochondrial O 2 l
in Mouse Aorta 3.3.1 Preparing Aortic Samples for HPLC Analysis (~3 h) Prepare fresh 2μM ...

Transfer 90μL of cell lysate to new Eppendorf tube containing 90 μL of 0.2 M HClO 4 , vortex, and keep on ice for 1 h to allow ...

CO 2 asphyxiation is a safe and humane method of euthanasia for mouse. It causes less oxidative stress, no observable histo- lo ...

E+ intercalates with DNA becoming highly fluorescent in nucleus. 2-OH-E+is majorly localized in cytosol. Therefore, 2-OH-E+ and ...

to evaluate oxidative stress using immunohistochemistry and Western blot [1, 18]. References Song P, Zou MH (2014) Redox regula ...

Chapter 33 Studying the Role of AMPK in Angiogenesis Katrin Spengler, Silke Große, Nderim Kryeziu, Anne Knierim, and Regine Hell ...

migration, and differentiation into tubular structures. Differentia- tion is best measured in three-dimensional assays, which in ...

1.2 Matrigel Plug Assay The Matrigel plug assay is a widely used in vivo assay, which allows the evaluation of angiogenesis with ...

and add a magnetic stirrer and autoclave (seeNote 5). Add 250 mL of prewarmed medium 199 (37C) to autoclaved methylcellulose, t ...

Reagents for protein determination (according to Lowry or Bradford). Multichannel pipettor for 100–200μL, sterile reservoir. St ...

Antidotes to anesthetics: 5 mg/mL atipamezole, 2.4 mg/mL naloxone, 1 mg/mL flumazenil. 1 mL syringes, 20- and 27-gauge needles. ...