AMPK Methods and Protocols

The luminometer measures the light produced for a period of 10 s. The plate is advanced to the next well for light measure- men ...

This step is required to allow cardiomyocytes to recover their basal state before to start hypertrophic treatment. 100 nM angio ...

protocol evaluates the final global incorporation of labeled phenylalanine into proteins. It integrates both entry and removal o ...

DAPI is a cell permeable fluorescent probe for DNA. It allows nuclear labeling. Once adenoviral construct is used, several bios ...

factor 2 kinase and to its phosphorylation at a novel site, serine 398. J Biol Chem 279 (13):12220–12231. https://doi.org/10. 10 ...

Chapter 22 Assessment of AMPK-Stimulated Cellular Long-Chain Fatty Acid and Glucose Uptake Joost J. F. P. Luiken, Dietbert Neuma ...

glucose does not readily cross the plasma membrane, and mem- brane transporters have long been recognized to mediate the bulk of ...

1.1 Considerations of the Uptake Assay 1.1.1 Applicability This protocol of measurement of cellular substrate uptake is dedi- ca ...

cardiomyocytes, this initial uptake phase amounts to 3–5 min after addition of the radiolabeled substrates (Fig.1a)[ 8]. In most ...

LCFA-albumin ratio roughly varies between 0.3 and 3. Of further note, possible changes in LCFA uptake would be most sensitively ...

the plasma membrane in clone-9 cells [18], GLUT1 activity in muscle cells is generally regarded as non-inducible, involved in me ...

2.3 Uptake Assay 1. Ethanol. DMSO. KOH: 1 M KOH. CaCl 2 : 100 mM CaCl 2. Bovine serum albumin (BSA)—Fraction V (fatty acid-free ...

Aliquot the supernatant into 2–5 mL portions inglassvials and store at 20 C. Solutions to Be Prepared Extemporaneously the Day ...

Spin the cells down for2 min at 4C(seeNote 9). Remove the supernatant and wash with 10 mL of ice-cold Stop solution. Repeat t ...

Table 1Data obtained from an example experimentCondition dpm 14 C Minus background Palmitate uptake Normalized dpm 3 H Minus bac ...

concentration is cytotoxic in cell lines, such as HL-1 cells, in which concentrations of up to 1μM are more advisable. Important ...

GLUT4 translocation in cardiomyocytes upon long-term (18 h) treatment, indicating reliance on synthesis of specific proteins. Th ...

Small-molecule direct AMPK activators: A screen by the Abbott Laboratories of >700,000 compounds yielded a non-nucleoside th ...

tissues, AMPKα2 knockout mice would be suitable, given that theα1 subunit is only marginally expressed in muscles. Fur- thermore ...

uptake into primary cardiomyocytes. AICAR-stimulated LCFA uptake was entirely inhibited, whereas contraction and oligomycin-stim ...