ATP is widely found in animals, plants, microorganisms and cultured cells, and is the currency of bioenergy, and energy charge is the main parameter describing the state of cellular energy metabolism. The ATP content is measured and the energy charge is calculated, which can reflect the state of energy metabolism.
Precautions: 1. If the supernatant after centrifugation is turbid, it is normal.
2. If the absorbance value is greater than 11. It is recommended that the sample be diluted with the extract for determination. Note that the calculation formula is multiplied by the dilution factor; If the absorbance value is too low.
or close to the blank, it is recommended to increase the sample size and carry out the measurement, and pay attention to the synchronous modification of the calculation formula.
Technical advantages:1The operation is simple, the empty plate is directly filled, and it can be detected after filling, and the time is short!
2.Suitable for precious samples: the entire experimental system only needs 20ul
3.Homogeneous detection: no coating, no washing ELISA, saving time and effort.
4.Stable system: It can be detected at any time within 7 days, and the signal is basically unchanged.
5.The data is objective and true: the ratio processing can effectively remove the background fluorescence, and the false positive rate and valence negative rate are low.
6.High-end detection, just a simple microplate reader can be easily done!
The ATP assay kit can be used to detect ATP (adenosine 5) in common solutions, cells, or tissues'-triphosphate) level. Cell and tissue samples can be prepared in a single lysed step, with detection sensitivitys up to 1 nmoll, chemiluminescence can be stable for up to 30 minutes, and samples can be simultaneously tested at the western level.
ATP, as an important energy molecule, plays an important role in various physiological and pathological processes of cells. Changes in ATP levels will affect the function of cells, usually cells will decrease in apoptosis, necrosis or in some toxic states, and high glucose stimulation can upregulate intracellular ATP levels for some cells. Usually a decrease in ATP levels indicates impaired or decreased mitochondrial function, and a decrease in ATP levels during apoptosis usually occurs at the same time as a decrease in mitochondrial membrane potential.
Sample preparation is very simple. The kit provides an ATP assay lysate that can be used for cell and tissue lysis and is ready for ATP detection after simple lysis. There is no need for perchloric acid or TCA extraction, or boiling after sample lysis.
Preparation of the solution:
1.Under the condition of low temperature of the extract, there may be crystallization precipitation, which can be heated and dissolved in a 60 water bath without affecting the use.
2.Reagent 2: Add 7 ml of distilled water to fully dissolve before use, which can be heated to promote dissolution;
3.Reagent 4: Take 1 stick and add 0Dissolve in 2ml of distilled water, and store the inexhaustible reagent-20 in aliquots for 2 weeks to avoid repeated freezing and thawing;
4.Reagent 5: Add 32 ml of distilled water is fully dissolved, and the inexhaustible reagent-20 is stored in aliquots for 4 weeks to avoid repeated freezing and thawing;
5.Reagent 6: Take 1 stick and add 025 ml of distilled water for later use, and the inexhaustible reagent-20 is stored in aliquots for 2 weeks to avoid repeated freezing and thawing;
6.Standard: 5 mg ATP. Add 0826 ml distilled water to prepare 10 mol of ATP standard solution, and store in unused reagent-20 aliquots for 4 weeks to avoid repeated freeze-thaw cycles;
7.Preparation of working solution: Before use, please press reagent two (ml): reagent three (ml): reagent four (ml): reagent five (ml): reagent six (ml) = 1:1:01：0.4：