Atp assay kit

ATP was measured through a colorimetric method by using an ATP assay kit (ElabScience, USA). Cell culture samples were prepared as follows: treated and untreated cell samples were collected and centrifuged at 1000–1500 r/min for 10 min. The supernatant was removed, and the cell sediment was retained (approximately 10⁶/ml cells). The cell suspension was prepared with 0.3–0.5 ml of boiled double-distilled water, allowed to stand in a boiling water bath for 10 min, blended, and finally extracted for 1 min. The sample was centrifuged at 3500 r/min for 10 min, and the supernatant was obtained for detection. The OD values of each tube were measured at 636 nm, following the manufacturer’s recommendation.

A431 cells with or without the indicated transfection were cultured normally for 24 h to collect the supernatant. The levels of glucose, lactic acid and ATP in the supernatant were analyzed with Glucose Assay kit (cat. no. E-BC-K234-S; Elabscience), L-Lactic Acid Colorimetric Assay kit (cat. no. E-BC-K044-S; Elabscience) and ATP Assay Kit (cat. no. S0026; Beyotime).

ATP contents were determined using a colorimetric method using an ATP assay kit (Elabscience, United States). A fresh mouse mammary adenocarcinoma tissue sample was weighed and cut into pieces, then added to boiled distilled water and incubated in a boiling water bath for 10 min, and then mixed fully for 1 min. The samples were centrifuged for 10 min at x10000 g. The supernatant was collected for measurement (according to the manufacturer’s instructions). The principle of the detection kit is that creatine kinase catalyzes creatine and adenosine triphosphate to generate creatine phosphate, then identified by phosphomolybdic acid colorimetry.