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Natural Science Forum / Biology / Biology / July 2004METHODS FOR STUDY ATP
Hi I will be much gratefull if somebody tell how can I measure ATP. I have use luciferin-luciferase assay but would like to compare this method with others. Kind Regards! Martin > I will be much gratefull if somebody tell how can I measure ATP. > I have use luciferin-luciferase assay but would like to compare this > method with others. You could use a coupled spectrophotometric assay, if sample volume and ATP-concentration are sufficiently high. ATP, like all nucleotides, has a strong absorbtion at 258 nm, which can be measured. If interfering substances are present, this can be done following ion exchange or ion pairing chromatography (see for example Ushimaru & Fukushima: Anal. Biochem 313 (2003) 173-5), using the area of the chromatographic peak as measured variable. Theoretically, it should also be possible to measure the concentration of ATP, like any other compound, by MS or (if there may be compounds of similar MW present) MS/MS. This would be a very sensitive method, if you can get hold of the instrument. > I will be much gratefull if somebody tell how can I measure ATP. > I have use luciferin-luciferase assay but would like to compare this > method with others. You could use a coupled spectrophotometric assay, if sample volume and ATP-concentration are sufficiently high. ATP, like all nucleotides, has a strong absorbtion at 258 nm, which can be measured. If interfering substances are present, this can be done following ion exchange or ion pairing chromatography (see for example Ushimaru & Fukushima: Anal. Biochem 313 (2003) 173-5), using the area of the chromatographic peak as measured variable. Theoretically, it should also be possible to measure the concentration of ATP, like any other compound, by MS or (if there may be compounds of similar MW present) MS/MS. This would be a very sensitive method, if you can get hold of the instrument. |
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