The enzymatic measurement of adenine nucleotides and P-creatine in picomole amounts
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Metabolic adaptation to the chronic loss of Ca<sup>2+</sup> signaling induced by KO of IP<inf>3</inf> receptors or the mitochondrial Ca<sup>2+</sup> uniporter
2022, Journal of Biological ChemistryCitation Excerpt :NADP+ and NADPH were measured by an enzymatic cycling assay using glucose-6-phosphate dehydrogenase (100). ATP was assayed in neutralized PCA extracts using a luciferase method (101). AMP was assayed using the AMP-glo kit (Promega) following the manufacturer's instructions.
Consequences of Neonatal Resuscitation with Supplemental Oxygen
2008, Seminars in PerinatologyCitation Excerpt :To determine the degree and duration of hypoxia necessary to demonstrate production of cortical lactate, rat pups were randomized to be ventilated with 21% or 5% O2 for up to 60 minutes. At each time point, beginning after 5 minutes exposure to 21% or 5% O2, a whole-brain freezing method of fixation16 with liquid nitrogen was performed and the frontal cortices of the pups were dissected and kept at −80°C. Lactate concentration in tissue, as a measure of cellular energy metabolism, was determined by standard luciferin–luciferase bioluminescence and biochemical assays, respectively, as previously reported.16
Adenosine treatment delays postischemic hippocampal CA1 loss after cardiac arrest and resuscitation in rats
2006, Brain ResearchCitation Excerpt :The samples of the cortex, brain stem and hippocampus were dissected from frozen lyophilized sections. Levels of ATP and total adenylates (AXP = ATP + ADP + AMP) were analyzed in tissue samples by microquantitative histochemistry assay methods described previously (Lust et al., 1981). Metabolite levels were reported as nanomoles per milligram dry tissue weight.
Compromised metabolic recovery following spontaneous spreading depression in the penumbra
2004, Brain ResearchCitation Excerpt :The tissue at the sites of the various electrodes was not sampled to avoid the potential artifact from the trauma of the craniectomy and/or insertion of the probe. The pieces of tissue were extracted in oil-well racks either in 0.02N HCl or 0.02N NaOH and the levels of adenosine triphosphate (ATP), P-creatine (phosphocreatine), glycogen, glucose, lactate, γ-aminobutyric acid (GABA), citrate and inorganic phosphate (Pi) were determined by enzymatic analysis [33,34]. The results from these experiments are presented graphically to show the differences between the metabolic consequences of SD in normal and ischemic brain.