BioLux Gaussia Luciferase Flex Assay Kit

• 产品编号：E3308S      品牌：New England Biolabs       原厂货号：E3308S
• 产品分类：蛋白质检测与分析 > 酶检测和活性分析 > 荧光素酶检测
• 应用分类：

描述：

特性：

■   启动子/增强子分析
■  高通量分析
■  转染条件优化
■  与其它报告系统一起进行多重分析
■  分泌途径分析
■  siRNA 筛选
■  时间曲线研究
■  单细胞分析，包括干细胞和原代细胞
■  活细胞分析

概述：NEB 为研究者提供可以检测细胞培养物上清或裂解物中 Gaussia 荧光素酶 (GLuc) 或 Cypridina 荧光素酶 (CLuc) 活性的试剂盒。因为 GLuc 和 CLuc 使用不同的底物，并且无交叉反应，因此，当一种荧光素酶存在时，并不影响另一种荧光素酶的活性测定，这使得这些荧光素酶可作为双重报告系统使用。
BioLux® Gaussia 荧光素酶检测试剂盒含有检测 GLuc 活性所必需的试剂。该试剂盒最常用于直接检测细胞上清液，也可用于检测细胞裂解液。通过精心设计，该试剂盒具有最大的光信号输出。BioLux Gaussia 荧光素酶检测试剂盒也可用来检测海肾荧光素酶的活性，因为 GLuc 和海肾荧光素酶都可以氧化腔肠(coelenterazine) 。
BioLux Gaussia 荧光素酶 Flex 检测试剂盒可以灵活选择较强的荧光信号或较长的信号半衰期。该试剂盒包括较高浓度的底物和一种稳定剂，加入后可以增加信号半衰期。该试剂盒是高通量筛选的理想工具 。
BioLux Cypridina 荧光素酶检测试剂盒含有检测 CLuc 活性所必需的试剂。该系统可用于检测细胞培养物上清或细胞裂解物。该系统使用 vargulin 荧光素作为底物。

BioLux Cypridina 荧光素酶启动试剂盒含有两个 CLuc 编码质粒，以及检测 CLuc 活性所需试剂。

原厂资料：

注意事项：

BioLux GLuc Flex Assay Buffer and Stabilizer can be stored at 4°C.

BioLux GLuc Flex Substrate must be tightly capped and stored at -20°C.

Usage notes:

1. Because of the stability of GLuc, the activity measured in the growth media of a GLuc-expressing culture reflects the protein that has accumulated up to the time of sampling.
2. The GLuc assay solution should be allowed to equilibrate at room temperature for 25 minutes (protect from light in a tightly capped tube/bottle) before adding to the sample.
3. After adding the equilibrated GLuc assay solution to the sample, we recommend a delay time of 35–40 seconds before taking a measurement, in order to reach the maximum level of detection. This is especially important when the GLuc activity level is low (e.g. < e4 RLU). For example, the readout obtained after 35–40 seconds of delay is ~e4; when compared to 30, 20 and 10 seconds of delay, the detection level is ~98% for 30 seconds of delay, ~93% for 20 seconds of delay & ~80% for 10 seconds of delay (Figure 6).
4. Use the prepared assay solution within 24 hours. The unused portion of the assay solution should be tightly capped and stored at -20°C. It should be completely thawed in the dark at room temperature before use.
5. The linear range of the luminometer used for the assay must be established. This is easily done by assaying serial dilutions of a sample. In addition, the assay solution itself, as well as the conditioned media (growth media from untransfected cells) should be included to establish the background in the assay.
6. If excess activity for the instrument range is found, the sample should be diluted in either PBS or 10% serum-containing media. The integration time can also be reduced (e.g. 2 seconds instead of 5 seconds).
7. When assaying the serial dilutions of a sample, it is best to assay the most diluted samples first & the most concentrated samples last. This will help to minimize false readings, i.e. cross talk effect in which signals from samples of high RLU cross into the next sample. The cross-talk effect seems to be more pronounced when white or black plates with clear-bottoms are used.