ATP/ADP/AMP Assay Kit

Price $300
Product Description: Protocol Download
Many metabolic reactions are controlled by the energy status of the cell. One index of the energy status is the energy charge (EC) of the cell, which is determined by the concentrations of adenine nucleotides. Although EC could theoretically range in values from 0 to 1, most cells maintain a steady-state EC in the range of 0.8 to 0.95, meaning that the energy status of the cell is buffered. The adenylate nucleotides ATP, ADP and AMP are also allosteric effectors for many enzymes. Intracellular adenine nucleotide concentrations vary during cell growth and cell death. Extracellular adenine nucleotides can regulate various signaling pathways. The assay kit is based on the luciferase reaction in the presence of ATP, which generates yellow-green light that can be conveniently measured by a luminometer or scintillation counter (Anal. Biochem. 80:496,1977). This provides the basis for a sensitive ATP assay system capable of detecting 1 picomole of ATP. The ATP/ADP/AMP Assay Kit is sufficient for 200 assays using 0.1 ml of ATP Assay Solution per assay. Sample ADP and AMP are converted to ATP by pyruvate kinase- and adenylate kinase-mediated reactions. Note that calculated sample AMP/ADP concentrations may be over- or underestimated due to the multiple factors that can affect nucleotide conversion in vitro. The ATP Assay Solution if stored in aliquots at -80°C is stable for many years. Repeated freeze-thaw cycles should be avoided.