CellROX™ Green Reagent, for oxidative stress detection

氧化应激检测用探针（绿色）

订购信息：

以上所有产品均为Life（Invitrogen）原装正品，原始包装为10x50 µg（10支50µg单个包装构成）。我司对C10444、C10422进行拆开销售，特惠价为2180元/支，单支50µl，现货销售，欢迎订购。

背景介绍：

氧化应激（Oxidative stress）的产生，起因于反应氧物质（ROS）生成与细胞清除ROS之间的不平衡。ROS在几种疾病的发展中发挥重要作用，包括炎症、动脉粥样硬化、衰老和衰老相关的退行性疾病。

基本介绍：

CellROX® Green Reagent是一种检测活细胞氧化应激的新型荧光探针。此细胞膜渗透性的染料在还原态时呈弱荧光，一旦被活性氧物质ROS氧化并结合到DNA后发明亮的绿色荧光，具有~485/520nm的最大激发/发射波长。此染料能够被甲醛固定且能承受去污剂处理，因此特别适合与其他兼容性染料和抗体进行多重标记检测。

CellROX® Green Reagent与各种检测平台兼容，比如传统荧光显微镜、高内涵筛选仪器（HCS）、流式细胞仪、荧光酶标仪或高通量筛选（HTS）。还与各种台式仪器兼容，比如FLoid™，Tali™，和Attune™仪器。

产品参数

1）外观：浅黄色溶液（pale yellow solution）

2）溶剂：DMSO

3）激发/发射光谱范围：可见光/可见光

4）亚细胞定位：线粒体、细胞核

5）保存方法：≤-20°C避光干燥

染色示例

Fig1. Nefazodone Induced Oxidative Stress Measured with CellROX™ Green Reagent in HepG2 Cells

Nefazodone induced oxidative stress in hepatocarcinoma (HepG2) cells measured with CellROX™ Green Reagent. HepG2 cells were plated on glass bottom 35 mm MatTek dishes. The cells were treated with or without 50 µM nefazodone for 24 hr at 37° C. The cells were then stained with 5 µM CellROX™ Orange Reagent and Hoechst 33342 by adding the probes to complete media and incubating at 37° C for 30 min. The cells were washed with PBS and imaged on a Zeiss Axiovert inverted microscope using a 40x objective. Nefazodone treatment leads to increased oxidative stress.

Fig2. MnTBAP Inhibition of Menadione Induced Oxidative Stress Measured with CellROX™ Green Reagent in BPAE Cells
MnTBAP inhibition of menadione induced oxidative stress in bovine pulmonary endothelial (BPAE) cells measured with CellROX™ Green Reagent. BPAE cells were plated on glass bottom 35 mm MatTek dishes. The cells were treated with or without 100 µM menadione for 1 hr at 37° C. 100 µM MnTBAP was added to the dish with or without menadione treatment. The cells were then stained with 5 µM CellROX™ Green Reagent and Hoechst 33342 by adding the probes to complete media and incubating at 37° C for 30 min. The cells were washed with PBS and imaged on a Zeiss Axiovert inverted microscope using a 40x objective. MnTABP co-incubation with menadione reduced the oxidative stress in the cells.

常见问题

1）I am trying to label my cells with a reactive oxygen species (ROS) indicator dye, but I am not seeing a significant difference in signal. What could be happening?

First, make sure you have both a negative (untreated) and positive (ROS-induced) sample to compare. A good positive control can be the use of 100 μM menadione for one hour or 50 μM nefazodone for 24 hours. H2O2 can also be used, though it does not work well for CellROX™ dyes. Some dyes, such as H2DCFDA, require esterase cleavage, so don't incubate in the presence of serum (which contains esterases that can prematurely cleave the dye). If your positive control does not show significant change compared to the negative control, try increasing the concentration and label time for the dye. Our manuals give starting recommendations. Be sure to image your live cells as soon as possible. Only two dyes (CellROX™ Green and CellROX™ Deep Red) are retained with formaldehyde fixation. Finally, make sure you are using filters and instrument settings to match the excitation and emission spectra of the dye.

2）I need a formaldehyde-fixable reactive oxygen species detection assay. Is H2DCFDA fixable?

H2DCFDA and similar derivatives are not fixable. The same goes for dihydroethidium and dihydrorhodamine. However, CellROX™ Deep Red and CellROX™ Green are retained for a limited time upon fixation with formaldehyde. CellROX™ Green may be retained upon subsequent Triton™ X-100 permeabilization. Avoid the use of any acetone or alcohol-based fixatives or fixatives that include alcohol, such as formalin.

 — —Written/Edited by V. Shallan【版权归MKBio懋康所有】

上海懋康生物科技有限公司是一家涉足于生命科学和生物技术领域研究的试剂、仪器和实验室消耗品与实验服务工作，主要从事细胞生物学、植物学、分子生物学、免疫学、生物化学、蛋白组学。生物制药与诊断试剂研发生产等领域。 本公司秉承“以人为本，以诚为信、合同守信”的经营理念。坚持"品质保障"的原则为广大客户提供优质产品。