CellROX? Green Reagent, for oxidative stress detection

氧化應激檢測用探針（綠色）

訂購信息：

以上所有產品均為Life（Invitrogen）原裝正品，原始包裝為10x50 μg（10支50μg單個包裝構成）。我司對C10444、C10422進行拆開銷售，特惠價為2180元/支，單支50μl，現貨銷售，歡迎訂購。

背景介紹：

氧化應激（Oxidative stress）的產生，起因于反應氧物質（ROS）生成與細胞清除ROS之間的不平衡。ROS在幾種疾病的發展中發揮重要作用，包括炎癥、動脈粥樣硬化、衰老和衰老相關的退行性疾病。

基本介紹：

CellROX® Green Reagent是一種檢測活細胞氧化應激的新型熒光探針。此細胞膜滲透性的染料在還原態時呈弱熒光，一旦被活性氧物質ROS氧化并結合到DNA后發明亮的綠色熒光，具有~485/520nm的最大激發/發射波長。此染料能夠被甲醛固定且能承受去污劑處理，因此特別適合與其他兼容性染料和抗體進行多重標記檢測。

CellROX® Green Reagent與各種檢測平臺兼容，比如傳統熒光顯微鏡、高內涵篩選儀器（HCS）、流式細胞儀、熒光酶標儀或高通量篩選（HTS）。還與各種臺式儀器兼容，比如FLoid?，Tali?，和Attune?儀器。

產品參數

1）外觀：淺黃色溶液（pale yellow solution）

2）溶劑：DMSO

3）激發/發射光譜范圍：可見光/可見光

4）亞細胞定位：線粒體、細胞核

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.

常見問題

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【版權歸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學和生物技術領域研究的試劑、儀器和實驗室消耗品與實驗服務工作，主要從事細胞生物學、植物學、分子生物學、免疫學、生物化學、蛋白組學。生物制藥與診斷試劑研發生產等領域。 本公司秉承“以人為本，以誠為信、合同守信”的經營理念。堅持"品質保障"的原則為廣大客戶提供優質產品。