Applications of ATP Assay Kits in Cell Research

1. Define the Core Question: For quantifying total cell viability, choose bioluminescence; to visualize dynamic changes of ATP within cells, choose fluorescent probe methods; for in-depth analysis of metabolic pathways, choose real-time rate assays.

2. Emphasize Standardization: Ensure consistent cell seeding numbers, precise reagent addition volumes, and reading within the specified time window (e.g., 10-30 minutes) to improve reproducibility.

3. Validate the Linear Range: Before formal experiments, it is recommended to test with gradient cell numbers to ensure the signal shows good linearity within the detection range.

Note: The following workflow is a general example; for specific operations, please strictly follow the official instructions of your specific kit.

1. Step 1: Plating and Treatment: Seed cells at appropriate density in white opaque 96-well plates; after overnight culture, apply different treatments (e.g., drug intervention, environmental stress, etc.).
2. Step 2: Equilibration and Reagent Addition: Equilibrate the culture plate and detection reagents at room temperature for 30 minutes. Carefully remove old medium, add fresh medium containing detection reagents (one-step method), or first add cell lysis buffer to lyse cells followed by detection reagents (two-step method), ensuring consistent volumes per well.
3. Step 3: Detection and Reading: Place the plate on a microplate shaker and mix for 30 seconds; incubate at room temperature protected from light for 10 minutes, then read the relative light units (RLU) for each well using a luminometer.
4. Step 4: Data Analysis

   • Cell Viability (%) = (Average RLU of treatment group / Average RLU of control group) × 100%

   • Plotting: Typically, plot cell number on the x-axis (log scale) and RLU on the y-axis to generate a standard curve to assess linearity; plot drug concentration on the x-axis and cell viability on the y-axis to generate dose-response curves for evaluating drug effects on cell viability.

The Absin ATP Assay Kit is a specific and sensitive kit for detecting adenosine triphosphate (ATP). In biological organisms, energy substrates are ultimately metabolized primarily to ATP for energy consumption in biological activities. By quantitatively detecting ATP, one can determine cell proliferation, cell damage, cell activation, and intracellular granule release.

This kit has been specially optimized to provide excellent detection linearity with higher accuracy than general kits. Additionally, it offers high temporal stability by slowing down light signal decay, allowing luminescence signals to be measured within 5-30 minutes after reaction initiation.

This kit detects ATP in the linear range of 0.001-1 μM, with sensitivity ≤ 0.001 μM.

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