ATP Luminescence Cell Viability Assay Kit, 10 mL

Product Information

Product Description/Introduction

ATP is an important energy molecule in the cell which plays an important role in various physiological and biochemical activities of the cell, and is known as the energy currency in the organism. The cellular state and cell numbers show a correlation with ATP. Therefore, cell number or viability can be assessed by the relative level of ATP.

This kit is based on the principle of ATP-dependent luciferase catalyzing luciferin to produce fluorescence, which can effectively detect and evaluate the intracellular ATP level. Compared with the traditional methods such as CCK-8 and MTT, fluorescence method has the advantages of high sensitivity and wide range.

Storage and Shipping Conditions

Ship with dry ice; Store at -80 away from light for 12 months; Store at -20°C away from light, recommended for use within 6 months.

Product Content

Assay Protocol / Procedures

1. Cell pre-processing work: Inoculate the cells at a certain density in a 96-well plate (please choose a white opaque well plate suitable for cell culture or use a regular cell culture plate. When performing the detection, transfer the solution to the white detection wells to minimize interference between adjacent wells.) and pre-treat the cells according to the purpose of the experiment;

2. Sample preparation for ATP standards (optional): The ATP standard (self-contained) is gradually diluted with buffer such as PBS or basal medium and added to the well plate at 100 μL/well;

3. Cell Viability Assay:

3.1. Take out the ATP Luminescence Cell Viability Assay Reagent in advance, thaw it and return it to room temperature;

3.2. Remove the cell culture plate and equilibrate at room temperature for 5-10 min;

3.3. The ATP Luminescence Cell Viability Assay Reagent is added directly to the well plate at 100 μL/well;

3.4. Shake horizontally for 1-2 min or tap to mix;

3.5. After standing for 3 min, the chemiluminescence assay can be performed using a Luminometer, a multifunctional enzyme labeler with a chemiluminescence detection module, or other instruments capable of detecting bioluminescence (note: the sample itself does not emit light in the fluorescence method, but needs to be excited by a specific wavelength of excitation light and then received through a special channel. In chemiluminescence, the sample itself emits light and does not need to be excited by a specific wavelength of excitation light in order to be detected by the corresponding equipment);

4. Data analysis: The following data analysis is based on subtracting the background blank

4.1. Relative cell number (proliferation) analysis: According to the fluorescence intensity value, determine the number of cells.

4.2. Cell viability calculation (cell proliferation viability or cytotoxicity viability):

Note:

A (dosing group): treated cells + ATP luminescence cell viability assay reagent

A1 (no dosing group): normal untreated cells + ATP luminescence cell viability assay reagent

A0 (blank control group): cell culture medium (no cells) + ATP luminescence cell viability assay reagent

4.3. Quantitative analysis of ATP content (optional):

4.3.1. The fluorescence intensity value of the detected ATP standard curve is subtracted from the fluorescence intensity value data of the blank control group, and the standard curve is plotted using software such as Excel, and the formula is obtained:

where a represents the slope and b represents the intercept.

4.3.2. The X-value (ATP content) in the assay system is calculated according to the formula:

Calculate the value of Y by substituting the slope a and intercept b from 4.3.1 into the above equation.

4.3.3. Calculation of ATP content in samples:

Note

1. To ensure that the test reagent is used effectively, it is recommended to store in small portions and avoid repeated freezing and thawing.

2. The testing time warp is controlled within 5-30 min to ensure the accuracy of the testing data.

3. Part of the reagent precipitation after thawing, is a normal phenomenon, before use fully shaken to ensure its complete dissolution.

4. Try to avoid light and add the test reagents within a short period of time. It is recommended to use a multiwell pipette to add samples and pay attention to the consistency of the pipetting volume in each well of the pipette.

5. For your health and safety, please wear lab coat and gloves during operation.

For Research Use Only!

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