Aposensor adp atp ratio assay kit

Manufactured by Enzo Life Sciences

Sourced in United States

The ApoSENSOR ADP/ATP Ratio Assay Kit is a quantitative colorimetric assay designed to measure the ratio of ADP to ATP in biological samples. The kit utilizes an enzyme-coupled reaction to produce a color change proportional to the ADP/ATP ratio.

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Lab products found in correlation

Fluoroskan ascent, by Thermo Fisher Scientific (1 mentions) Fluostar optima spectrophotometer, by BMG Labtech (1 mentions)

2 protocols using aposensor adp atp ratio assay kit

Quantifying ADP/ATP Ratio in Apoptosis

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ADP/ATP ratios were measured at time points in which the maximum percentage of apoptotic cells and highest activity of caspase-3/7 were detected. The ApoSENSOR ADP/ATP Ratio Assay Kit (Enzo Life Sciences, Farmingdale, NY, USA) was used according to the directions of the manufacturer. Supernatants from cells cultured in 96-well plates were collected for subsequent HMGB1 measurement (see below), whereas the adherent cells were treated with 60 μL of nucleotide-releasing buffer, and the plate was incubated at RT for 5 min. To quantify intracellular ATP, 100 μL of ATP monitoring enzyme was mixed with 50 μL of cell extract in a 96-well plate. The bioluminescent signal was recorded immediately using a Fluoroskan Ascent (Thermo, Waltham, MA, USA) luminometer. After adding 10 μL of ADP-converting enzyme, intracellular ADP was measured. The ADP/ATP values are presented.

Chavez-Dominguez R.L., Perez-Medina M.A., Lopez-Gonzalez J.S., Galicia-Velasco M., Matias-Florentino M., Avila-Rios S., Rumbo-Nava U., Salgado-Aguayo A., Gonzalez-Gonzalez C, & Aguilar-Cazares D. (2021). Role of HMGB1 in Cisplatin-Persistent Lung Adenocarcinoma Cell Lines. Frontiers in Oncology, 11, 750677.

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Zebrafish ADP/ATP Ratio Assay

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Measurements were taken using the ApoSENSOR™ ADP/ATP Ratio Assay Kit (Enzo® Life Sciences, Inc., Farmingdale, NY, USA). Methods were adapted for live zebrafish embryos from the kit instruction manual and Lu et al. (2011) (link). All luminescence readings were measured with a FLUOstar® Optima spectrophotometer (BMG Labtech, Cary, NC, USA). Initial trials were performed to ensure that the ERSE solution and embryos alone were not auto-luminescent. A total of 2 replicate plates were completed per time point, with 5 wells per treatment in each plate (total n=10). Predicted values for additive interactions between hypoxia and ERSE in the H+E group were calculated by subtracting the observed control values for ATP content from both the observed hypoxia and ERSE values, then adding those together (i.e. (HYP-CON)+(ERSE-CON)=Expected). This provides us with a baseline mathematical expectation of additivity for comparisons to observed H+E values, although may not fully reflect biological additivity.

Lindberg C.D, & Di Giulio R.T. (2019). Polycyclic Aromatic Hydrocarbon and Hypoxia Exposures Result in Mitochondrial Dysfunction in Zebrafish. Aquatic toxicology (Amsterdam, Netherlands), 216, 105298.

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