CheckLite ATP Eliminating Kit

Extraction procedure

Preparation of reagent

• ATP eliminating reagent is kept under vacuum in a reddish pink-labeled vial.
• Pour the reconstitution buffer from the yellow-labeled vial into the opened reddish pink-labeled vial and leave it at room temperature for a few minutes.
• Stir the vial gently so as not to produce foam until the contents are completely dissolved.
• Do not touch the rim of the vial or the top of the rubber plug directly with your hands, since this will sometimes raise the blank value of the reagent.
• One vial of ATP eliminating reagent can be used for more than 50 assays under normal condition.

Notes for handling samples

• The concentration of ATP in living cells changes rapidly. Therefore, extract ATP from cells or freeze the cells immediately after sampling. Even in the case of extraction, the inhibition or destruction of ATP-degrading enzymes may be necessary to prevent enzymes from decreasing the concentration of the ATP.

Pretreatment of samples

• Solid samples:
• Treat the sample with a stomacher or a homogenizer and then use the supernatant for the following measurement.
• Liquid samples:
• When the sample solution is turbid, colored, or contains inhibitory ions, such as Cl-, dilute the sample as necessary.

ATP degradation of samples

• Put 1 ml of sample into the test tube.
• Add 0.1 ml of ATP eliminating reagent into the test tube and mix well.
• Let the test tube stand at room temperature for 30 to 40 minutes. It is important to keep the reaction time constant to obtain good reproducibility. If testing multiple samples, each test tube should stand at room temperature for the same time.
• After reaction at room temperature above, the samples are used for the bioluminescence assay below.