ATP 检测试剂，生物发光

用萤火虫荧光素酶检测活的哺乳动物细胞中的ATP

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产品号 #100-1542_C

用萤火虫荧光素酶检测活的哺乳动物细胞中的ATP

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总览

ATP检测试剂，生物发光使用一步法，仅需20分钟即可测定ATP，以监测在细胞增殖、细胞毒性和其他基于细胞实验中存活的哺乳动物细胞的数量。该实验使用萤火虫荧光素酶，这是一种61 kDa的单体酶，负责催化萤火虫荧光素的氧化。该反应产生稳定且持久（> 2 h）的560 nm光发射，直接与细胞中的 ATP 浓度相对应。由于萤火虫荧光素酶不需要二硫苏糖醇（DTT）的酶稳定作用，因此这种检测方法可以可靠地检测每孔中少至 50 个细胞，而且没有难闻的气味。ATP检测试剂，生物发光是即用型单瓶试剂（无需清洗细胞），可用于 96 孔或 384 孔微孔板，是高通量筛选实验的理想解决方案。可使用配备检测荧光素发光功能（luminometer）的酶标仪进行定量。

实验数据

Luminescent Signal Output vs. Cells Number Graph

Figure 1. ATP Assay, Bioluminescence Shows Luminescent Signal Output Is Directly Proportional to Cell Number

(A) HEK293 cells were plated in a white opaque 96-well plate with two-fold serial dilution at a density of 50 - 50 x 10^3 cells per well. Luminescent output was measured 20 minutes after addition of ATP Assay, Bioluminescence and shows a linear relationship (r2 = 0.99) between cell number and luminescence signal over three orders of magnitude and can detect as low as 50 cells per well. Values are representative of three replicates per cell number. (B) ATP Assay, Bioluminescence shows a linear relationship (r2 = 0.99) between cell number per well and luminescence signal even at low seeding densities (100 - 1600 cells).

Figure 2. ATP Assay, Bioluminescence has a Stable Luminescent Signal with a Half-Life of More than 2 Hours

HEK293 cells were plated in a white opaque 96-well plate at a density of 25 x 10^3 cells per well. After addition of ATP Assay, Bioluminescence, plate was incubated at room temperature. Luminescent output was measured 20 minutes after reagent addition and additional luminescence measurements were recorded at 60, 120, 180, 240, and 300 minutes. The graph shows ATP Assay, Bioluminescence half-life of the luminescent signal is more than 2 hours.

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ATP Assay, Bioluminescence

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