产品介绍
1. 外观:黄色粉末
2. 纯度:≥99% (HPLC)
3. 产品描述
D-虫荧光素(D-Luciferin)是萤火虫荧光素酶底物,其量子效率为0.88,是Luminol的20倍。反应原理:首先,在镁离子存在下荧光素酶使荧光素与ATP反应,接着它被氧化形成二氧杂环丁烷结构并发出黄绿色的光。Luciferin-luciferase发光用于ATP监控以测定细胞活力以及细菌计数。它还用于报告基因检测。可与小动物活体成像系统配套使用,用于标记LUC基因后的体内活体荧光检测。
4. 应用
(1) 体外分析
(2) 活体成像分析
(3) 高灵敏度ATP分析
使用方法
1. 活体成像分析方法
(1) 用DPBS(w/o Mg2+、Ca2+)配置D-Luciferin工作液(15mg/mL)* ,0.2 μm滤膜无菌过滤。
(2) 注射量:10μL/g的体重,如10g 重小鼠,注射100μL 工作液(1.5 mg D-Luciferin)。
(3)腹腔注射10~15分钟后,上机进行图像分析。
2. 体外分析方法
(1)用无菌水配置200× 的D-Luciferin储备液(30mg/mL)*,立即使用,或-20℃保存。
(2) 用预热好的完全培养基1∶200稀释D-Luciferin储备液,配置工作液(150μg/mL)。
(3)去除培养细胞的培养基。
(4)进行图像分析前,向细胞中添加1× 的D-Luciferin工作液,进行图像分析**。
* :溶解D-Luciferin时要完全。
**:在图像分析前,将细胞进行37℃的短时间培养可以增加信号强度。
相关产品
D-虫荧光素游离酸(D-Luciferin, free acid)、D-虫荧光素钾盐(D-Luciferin, potassium salt)和D-虫荧光素钠盐(D-Luciferin, sodium salt),钾盐、钠盐的形式是最通用的,因为它们都易溶于水。钾盐也是活体动物检测使用的主要形式。它们的激发和发射波长分别为328nm和533nm。
储存条件:-20℃干燥避光保存,有效期一年。
Product introduction
1. Appearance: yellow powder
2. Purity: ≥99% (HPLC)
3. Product description
D-Luciferin is a firefly luciferase substrate with a quantum efficiency of 0.88, which is 20 times that of Luminol. Reaction principle: First, luciferase reacts with ATP in the presence of magnesium ions, and then it is oxidized to form a dioxetane structure and emit yellow-green light. Luciferin-luciferase luminescence is used for ATP monitoring to determine cell viability and bacterial count. It is also used for reporter gene testing. It can be used with small animal live imaging system for in vivo fluorescence detection after labeling LUC gene.
4. Application
(1) In vitro analysis
(2) In vivo imaging analysis
(3) Highly sensitive ATP analysis
Instructions
1. In vivo imaging analysis method
(1) Prepare D-Luciferin working solution (15mg/mL)* with DPBS (w/o Mg2+, Ca2+), and filter aseptically with 0.2 μm filter.
(2) Injection volume: 10μL/g body weight, such as 10g heavy mouse, 100μL working solution (1.5 mg D-Luciferin) is injected.
(3) After 10-15 minutes of intraperitoneal injection, go to the computer for image analysis.
2. In vitro analysis methods
(1) Prepare 200× D-Luciferin stock solution (30mg/mL)* with sterile water, use it immediately, or store at -20°C.
(2) Dilute the D-Luciferin stock solution with the pre-warmed complete medium 1:200 and prepare the working solution (150μg/mL).
(3) Remove the medium for culturing the cells.
(4) Before image analysis, add 1× D-Luciferin working solution to the cells for image analysis**.
*: Dissolve D-Luciferin completely.
**: Before image analysis, cells are incubated for a short time at 37°C to increase the signal intensity.
Related Products
D-Luciferin free acid (D-Luciferin, free acid), D-Luciferin potassium salt (D-Luciferin, potassium salt) and D-Luciferin sodium salt (D-Luciferin, sodium salt), potassium salt The sodium salt form is the most versatile, because they are easily soluble in water. Potassium is also the main form used in live animal testing. Their excitation and emission wavelengths are 328nm and 533nm, respectively.
Storage conditions : Store at -20°C dry and dark, valid for one year.