CellROX™ Green Reagent, for oxidative stress detection

氧化應激檢測用探針（綠色）

訂購信息：

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

背景介紹：

氧化應激（Oxidative stress）的産生(shēng)，起因于反應氧物(wù)質（ROS）生(shēng)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，包括炎症、動脈粥樣硬化、衰老和衰老相(xiàng)關的退行性疾病。

基本介紹：

CellROX® Green Reagent是一(yī)種檢測活細胞氧化應激的新型熒光(guāng)探針。此細胞膜滲透性的染料在還(hái)原态時呈弱熒光(guāng)，一(yī)旦被活性氧物(wù)質ROS氧化并結合到(dào)DNA後發明亮的綠色熒光(guāng)，具有~485/520nm的最大激發/發射波長(cháng)。此染料能(néng)夠被甲醛固定且能(néng)承受去污劑處理，因此特别适合與其他兼容性染料和抗體進行多(duō)重标記檢測。

CellROX® Green Reagent與各種檢測平台兼容，比如傳統熒光(guāng)顯微鏡、高(gāo)内涵篩選儀器(qì)（HCS）、流式細胞儀、熒光(guāng)酶标儀或高(gāo)通(tōng)量篩選（HTS）。還(hái)與各種台式儀器(qì)兼容，比如FLoid™，Tali™，和Attune™儀器(qì)。

産品參數

1）外觀：淺黃色溶液（pale yellow solution）

2）溶劑：DMSO

3）激發/發射光(guāng)譜範圍：可見(jiàn)光(guāng)/可見(jiàn)光(guāng)

4）亞細胞定位：線粒體、細胞核

5）保存方法：≤-20°C避光(guāng)幹燥

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(jiàn)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】

上海夢澤生物科技有限公司是一(yī)家涉足于生(shēng)命科學和生(shēng)物(wù)技(jì)術(shù)領域研究的試劑、儀器(qì)和實驗室消耗品與實驗服務工(gōng)作，主要從(cóng)事(shì)細胞生(shēng)物(wù)學、植物(wù)學、分子生(shēng)物(wù)學、免疫學、生(shēng)物(wù)化學、蛋白(bái)組學。生(shēng)物(wù)制藥與診斷試劑研發生(shēng)産等領域。 本公司秉承“以人為(wèi)本，以誠為(wèi)信、合同守信”的經營理念。堅持"品質保障"的原則為(wèi)廣大客戶提供優質産品。