ATP测定，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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概述

ATP测定，生物发光是一个一步，20分钟的测定，测量ATP，以监测在细胞增殖，细胞毒性和其他基于细胞的测定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，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（bbb2小时）的560nm光发射，这直接对应于细胞中存在的ATP浓度。这种检测方法可以可靠地检测到每孔50个细胞，并且没有难闻的气味，因为萤火虫荧光素酶不需要用二硫苏糖醇（DTT）进行酶稳定。ATP Assay， Bioluminescence是一种单一的，即用型试剂（不需要洗涤细胞），可以在96孔或384孔微孔板中使用，使其成为高通量筛选实验的理想解决方案。定量可以使用能够发光的微孔板读取器（光度计）进行。

Data Figures

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.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

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Product Information Sheet

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ATP Assay, Bioluminescence

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