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            Thermo(invitrogen)單線態氧熒光探針 Singlet Oxygen Sensor Green(SOSG)
            時間:2018-12-26     作者:懋康原創     文章來源:懋康生物    

            單線態氧熒光探針 Singlet Oxygen Sensor Green(SOSG)



            產品關鍵詞:

            Singlet Oxygen Sensor Green®, SOSG;單線態氧(Singlet Oxygen,1O2);單線態氧熒光探針;S36002;Thermo Fisher; ROS(Reactive oxygen species)活性氧物質;

             

            活性氧引起的許多生理損傷都可能歸因于單線態氧(1O2),包括與脫氧鳥苷選擇性反應形成8-羥基脫氧鳥苷(8-OHdG)的核酸修飾。單線態氧的壽命足夠長(水中有4.4μs)使其能夠充分擴散進入細胞和組織。實驗室中通常用三種之一的方法來生成單線態氧:1)使用感光染料對二氧(3O2)進行光化學反應來制備;2)通過對一種過氧化物或二氧環丁烷進行加熱分解制備;3)氧氣流內通過微波放電來制備;單線態氧通過在1270nm處具弱化學發光的典型特征直接被檢測。

             

            Thermo fisher(molecular probe)公司開發的Singlet Oxygen Sensor Green®, SOSG是目前商業化可得且應用最廣泛的單線態氧熒光探針,從材料學到藥物學領域,比如光動力學療法。

             

            【特異性強】

            Singlet Oxygen Sensor Green®, SOSG高度選擇性結合單線態氧(1O2)。不像其他的熒光和化學發光的單線態氧檢測試劑,SOSG與其他活性氧物質(ROS),包括羥基自由基(·OH)、超氧陰離子(·O2–)和一氧化氮(NO)無可見反應【見圖1】。

            圖1、Singlet Oxygen Sensor Green (SOSG)的熒光反應和對單線態氧(1O2)的特異性。A)用熒光光度計(Ex/Em= 488/525nm)測定溶液內的熒光,溶液內含SOSG(1 μM )和亞甲基藍(10 μM)溶于100mM pH 7.5 Tris Buffer; 單線態氧清除劑疊氮鈉(NaN3,1 mM);或50% D2O(重水,能夠延長1O2的壽命)。每20s完成一次熒光測定,每次測定間隔30s(如灰條標示)。在這30s的間隔中,樣本暴露于激光輻射(630–680 nm,<5 mW),導致亞甲基藍光敏生成1O2。B)用熒光光度計(Ex/Em=488/525nm)測定溶液內的熒光,溶液為50mM pH 7 Tris buffer,1mM 黃嘌呤,SOSG(1 μM ),或含SOSG(1 μM ),或含二氫羅丹明123。約20s之后,加入50 mU/mL黃嘌呤氧化酶(XO)。XO催化黃嘌呤的氧化,產生尿酸和超氧化物。超氧化物瞬間被H2O2降解。

             

            【熒光特征】

            Singlet Oxygen Sensor Green®, SOSG與單線態氧(1O2)反應前,探針最初發弱藍色熒光,激發峰在372nm和393nm,發射峰在395nm和416nm。當含單線態氧(1O2),探針發射綠色熒光,光譜特征類似于熒光素(最大激發/發射波長約504/525nm)。

             

            【線性化關系】

            研究發現Singlet Oxygen Sensor Green®, SOSG的熒光產物在某些溶液中隨著時間被降解,且在不含單線態氧的堿性pH溶液中,Singlet Oxygen Sensor Green®, SOSG會發熒光。不過,當進行適當的操作,綠色熒光信號的強度與單線態氧(1O2)含量相關,且不會受其他ROS的明顯干擾。

             

            【應用】

            Singlet Oxygen Sensor Green®, SOSG證實能用來檢測溶液和植物組織中的單線態氧(1O2)。

             

            【使用方法】

            1.       SOSG儲存液制備:收到貨按照≤-20℃避光干燥保存。用甲醇來制備儲存液,每100μg粉末加入33μl甲醇制備~5mM儲存液。正式實驗前制備工作液,不能用完的工作液需丟棄,不可保存后再用。

            2.       溶液檢測:SOSG不具細胞膜滲透性,設計用來檢測溶液環境內的單線態氧。最佳的稀釋緩沖液和工作濃度需根據經驗來調整。建議起始工作濃度為1-10μM。

             

            【使用方法-來自文獻】

            一、文獻來源:Flors C et al. Imaging the production of singlet oxygen in vivo using a new fluorescent sensor, Singlet Oxygen Sensor Green. J Exp Bot. 2006;57(8):1725-34. Epub 2006 Apr 4.

            圖2. 肌紅蛋白加入脂質體后隨著時間的SOSG熒光變化(Changes in SOSG fluorescence with time after the addition of myoglobin to liposomes)。Singlet oxygen sensor green (SOSG) reagent was dissolved in methanol to make a stock solution of 500 μM. SOSG (final concentration 50 μM) was added to liposomes (2 mg ml?1) in 50 mM sodium phosphate buffer pH 7.4 in a Petri dish (2 cm diameter). Lipid oxidation was started by the addition of ferric horse myoglobin (10 μM). Following excitation of SOSG at 480 nm using a 75 W xenon lamp (Leica UK Ltd, Milton Keynes, UK) with a 480 nm interference filter and heat filter, images of the fluorescence from SOSG in the samples were produced using a Peltier-cooled (200 K) charge-coupled device (CCD) camera (576×384 pixels, Wright Instruments, Enfield, UK) with a 532 nm interference filter in the objective.

            圖3. 擬南芥葉子內SOSG熒光產生的動力學變化(Kinetics of the development of SOSG fluorescence in Arabidopsis leaves)。Leaves were grown at a PPFD of 200 μmol m?2 s?1, detached and infiltrated with SOSG, and then the top halves of the leaves exposed to a PPFD of 650 μmol m?2 s?1 (high light). The bottom halves were kept in the dark; images were taken after 0, 5, 10, 20, 30, and 50 min.

             

            二、文獻來源:Chen T et al. Singlet Oxygen Plays an Essential Role in the Root’s Response to Osmotic Stress. Plant Physiol. 2018 Aug;177(4):1717-1727.

            圖4. PEG處理后單線態氧和碘化丙啶細胞滲透性的變化圖(Appearance of singlet oxygen and changes in cell permeability to propidium iodide after PEG treatment)。Root tips of 7-d-old seedlings stained with propidium iodide (PI) and singlet oxygen sensor green (SOSG). Median confocal slices are shown for each of these stains, along with the corresponding bright field (BF) image. Treatments were for the times indicated on the right: no treatment; 1 to 2 h, early; 3 to 4 h, mid; 5 to 8 h, late. Zones are as follows: AM, apical meristem; BM, basal meristem; and EZ, elongation zone. The median plane of the root was imaged where opaque regions in PI staining are indicative of cell death. White outlined arrows indicate SOSG and PI staining of live cells (i.e. the stain is observed in the perimeter of the cell). Solid white arrows indicate SOSG and PI staining of dead cells (i.e. where the PI stain permeates and fills the cell). Graphs on the left show quantification of the fluorescence, calculated by estimating the area of the root (using threshold above background) in a median slice that displayed SOSG or opaque PI fluorescence (using threshold above that obtained from live cells). Scale bar, 100 μm. For quantification, error bars are se, n ≥ 8.

             

            【爭議點】

            Singlet Oxygen Sensor Green®, SOSG不具有細胞膜滲透性,能用于細胞單線態氧(1O2)檢測?

            專利持有方thermo(molecular probe)官方說明此探針不具有細胞膜滲透性,適用于溶液體系單線態氧(1O2)檢測。然而,Gollmer A et al.報道SOSG能插入活哺乳動物細胞(文獻來源:Gollmer A et al. Singlet Oxygen Sensor Green®: photochemical behavior in solution and in a mammalian cell. Photochem Photobiol. 2011 May-Jun;87(3):671-9.)。

             

            訂購信息:Thermo fisher原裝正品,大包裝拆開單賣,100μg /支,單支特價700元。懋康生物長期現貨供應,歡迎咨詢,QQ:2971634497,或電話:021-54736159。

            品牌

            貨號

            名稱

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            特價(元)

            Thermo fisher

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            S36002

            Singlet Oxygen Sensor Green 單線態氧熒光探針

            100μg

            700

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            【溫馨提示】:見我司懋康生物整理的活性氧檢測探針專題(Probes for reactive oxygen species)。

             

            相關產品:ROS研究常用的配套產品,需要了解更多信息歡迎來電咨詢,021-54736159,QQ:2971634497。

            貨號

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            規格             

            價格(元)  

            MX4801-1KIT

            Reactive Oxygen Species (ROS) Assay Kit

            活性氧(ROS)檢測試劑盒

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            注意事項

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