Description

Applications

ATP Kit SL is used to assay both enzymes and metabolites. Any reaction possible to couple ATP formation or degradation e.g. kinases and their substrates or mitochondrial ATP production is possible to assay with the ATP Kit SL. ATP Kit SL contains ATP Reagent SL, a reagent emitting a stable light with a decay of less than 1%/min. The stable light makes it possible to contionously monitor the ATP levels in a sample. At the end of the assay the measurement is calibrated with BioThemas internal ATP Standard. Both kinetic and end-point assays can be performed using the 144-041 ATP Kit SL. ATP Kit SL is intended for monitoring of ATP (adenosine triphosphate) over the range of 10 ^–12 to 10 ^–6 mol/L.. The low decay rate of the light emission (around 0.9%/min) is due to a low consumption of ATP in the firefly luciferase reaction and a luciferase activity that is unchanged during the time of the measurement. Under these circumstances measurement of the intensity of the light emission allows continuous monitoring of formation or degradation of ATP in enzymatic reactions or release of ATP from cells. Thus assays can be performed in much the same way as NAD(P)H is spectrophotometrically or fluorometrically monitored in assays based on dehydrogenase reactions. Furthermore the stable light makes assays even of a constant ATP level more convenient and reliable, particularly when using manual luminometers.

If a higher sensitivity is required BioThema has two other reagents allowing the detection of 10 ^-17 mol and 10 ^-18, resp.

If no immediate reaction can be coupled to the metabolism of ATP, e.g. pyrosequencing or measurement of DNA polymerase, please contact us at info(at)biothema.com for recommendations of assay development.

For ATP determinations in eukaryotic cells we recommend Cell Viability Kit SL or Cellular ATP Kit HTS.

For ATP determinations in prokaryotic cells we recommend ATP Biomass Kit HS, Intracellular ATP Kit HS or Microbial ATP Kit HS, which all contain a more sensitive ATP reagent.

Instruments

The stable light makes it possible to use manual single tube luminometers, automatic tube luminometers or microplate luminometers. The detection limit obviously depends on the luminometer. With most luminometers 10 ^-15 moles of ATP can be detected.Dispensers are recommended for ATP Reagent SL and, in particular, for ATP standard.

Assay procedure

Detailed described procedures are found in the “Instructions for Use”, which is included with every kit.

Product characteristics

No. of determinations (cuvettes): 200No. of determinations (microplate): 1000

Detection limit:

Quantitative detection of ATP in the range of 10 ^–12 to 10 ^–6 mol/L.

Depending of the assay format this means 10 ^–15 to 10 ^–10 mol per assay (1 ml cuvette volume) for tube luminometers and 0.2_*10 ^–15 to 0.2 _*10 ^–10 mol per assay (0.2 ml volume) for microplate luminometers.