CellROX Deep Red Reagent, for oxidative stress detection

氧化應(yīng)激檢測(cè)用探針（深紅色）

溫馨提示：

見我司（懋康生物）整理的活性氧ROS熒光探針專題。

見我司（懋康生物）整理的CellROX Reagent, for oxidative stress detection氧化應(yīng)激檢測(cè)探針產(chǎn)品專題。

訂購(gòu)信息：

背景介紹：

氧化應(yīng)激（Oxidative stress）的產(chǎn)生，起因于反應(yīng)氧物質(zhì)（ROS）生成與細(xì)胞清除ROS之間的不平衡。ROS在幾種疾病的發(fā)展中發(fā)揮重要作用，包括炎癥、動(dòng)脈粥樣硬化、衰老和衰老相關(guān)的退行性疾病。

基本介紹：

CellROX Deep Red Reagent是一種檢測(cè)活細(xì)胞氧化應(yīng)激的新型熒光探針。此細(xì)胞膜滲透性的染料在還原態(tài)時(shí)無熒光或呈微弱熒光，一旦被活性氧物質(zhì)（ROS）氧化后發(fā)明亮穩(wěn)定的熒光，最大吸收/發(fā)射波長(zhǎng)約~ 644/ 665nm（見圖I.CellROX Deep Red氧化后的熒光光譜圖）。其熒光信號(hào)主要定位在細(xì)胞質(zhì)。

CellROX Deep Red Reagent的深紅色熒光信號(hào)與其他活細(xì)胞染料和GFP兼容，使其適用于多重?zé)晒庠囼?yàn)來測(cè)定各種細(xì)胞現(xiàn)象，包括與細(xì)胞毒性和細(xì)胞死亡的參數(shù)。不同于其它的ROS探針，比如H2DCFDA（MX4802-50MG），CellROX Deep Red Reagent的熒光信號(hào)經(jīng)醛類固定后也能保留，使其相對(duì)于傳統(tǒng)的ROS探針來說在應(yīng)用和操作流程更加靈活。CellROX Deep Red Reagent與各種檢測(cè)平臺(tái)兼容，比如傳統(tǒng)熒光顯微鏡、高內(nèi)涵篩選儀器（HCS）、流式細(xì)胞儀、熒光酶標(biāo)儀或高通量篩選（HTS）。還與各種臺(tái)式儀器兼容，比如Attune™儀器。

產(chǎn)品參數(shù)

1. 外觀：無色溶液（Colorless solution）

2. 溶劑：DMSO

3. 激發(fā)/發(fā)射光譜范圍：644/665nm（可見光/可見光）

4. 亞細(xì)胞定位：細(xì)胞質(zhì)

5. 熒光顏色：遠(yuǎn)紅（Far-red）

6. 保存方法：≤-20°C避光干燥

染色示例

Fig1. BPAE detection of oxidative stress using MitoTracker® Green (Cat. No. M7514) and CellROX® Deep Red Reagent (Cat. No. C10422) BPAE cells were plated in a 96 well plate, treated with 100 µM menadione for 1 hr and then stained with 20 nM MitoTracker® Green, 5 µM Far Red ROS Sensor and Hoechst 33342 for 30 mins in complete medium. Cells were then washed 3X with PBS and imaged on a Thermo Fisher Cellomics ArrayScan® VTI. The control sample has no signal while in menadione treated cells, there is a robust increase in the signal as a result of oxidative stress caused by menadione. Both MitoTracker® Green and Far Red ROS Sensor stained well showing that they work together in a multiplex assay.

Fig2. A549 spheroid mounted with Prolong Glass and imaged with EVOS FL Auto 2. A549 spheroid grown in Nunclon Sphera microplates (Cat. No. 174925) at 1000 cells/well. Live cells were labeled with NucBlue Live ReadyProbes Reagent (Cat. No. R37605), MitoTracker Orange (Cat. No. M7510), and CellROX Deep Red Reagent for oxidative stress detection (Cat. No. C10422) for 1 hour at 37°C. Cells were then fixed with 4% PFA, transferred to a #1.5 coverslip, and mounted in ProLong Glass. Sample was allowed to cure overnight exposed to air, and the coverslip was mounted to the slide using 100% glycerol. Image acquired with EVOS FL Auto 2 (AMAFD2000) using an Olympus 10X air objective (AMEP4753).

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.

3) What dyes can I use to detect reactive oxygen species (ROS) in my bacteria?

Many dyes that are used on mammalian cells have also been shown to be useful in bacterial cells. For example, CellROX Deep Red Reagent has been shown to work in B. subtilis (see Reference:bochum.de/netahtml/HSS/Diss/RaatschenNadja/diss.pdf). If you are interested in a particular dye, but are not sure if it will work on your bacteria, literature searches are the best way to check to see if it has been tested. If not, then it may be worth testing yourself.

注意事項(xiàng)

為了您的安全和健康，請(qǐng)穿實(shí)驗(yàn)服并戴一次性手套操作。

— —Written/Edited by V. Shallan【版權(quán)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(xué)和生物技術(shù)領(lǐng)域研究的試劑、儀器和實(shí)驗(yàn)室消耗品與實(shí)驗(yàn)服務(wù)工作，主要從事細(xì)胞生物學(xué)、植物學(xué)、分子生物學(xué)、免疫學(xué)、生物化學(xué)、蛋白組學(xué)。生物制藥與診斷試劑研發(fā)生產(chǎn)等領(lǐng)域。 本公司秉承“以人為本，以誠(chéng)為信、合同守信”的經(jīng)營(yíng)理念。堅(jiān)持"品質(zhì)保障"的原則為廣大客戶提供優(yōu)質(zhì)產(chǎn)品。

氧化應(yīng)激檢測(cè)用探針（深紅色）細(xì)胞染色 氧化應(yīng)激檢測(cè)用探針（深紅色）細(xì)胞染色