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    当前位置:首页> 新闻资讯> 新品发布 >Thermo(invitrogen)单线态氧荧光探针 Singlet Oxygen Sensor Green(SOSG)
    Thermo(invitrogen)单线态氧荧光探针 Singlet Oxygen Sensor Green(SOSG)
    时间:2018-12-26     作者:懋康原创     文章来源:懋康生物    

    单线态氧荧光探针 Singlet Oxygen Sensor Green(SOSG)



    产品关键词:

    Singlet Oxygen Sensor Green®, SOSG;单线态氧(Singlet Oxygen,1O2);单线态氧荧光探针;S36002;Thermo Fisher; ROS(Reactive oxygen species)活性氧物质;

     

    活性氧引起的许多生理损伤都可能归因于单线态氧(1O2),包括与脱氧鸟苷选择性反应形成8-羟基脱氧鸟苷(8-OHdG)的核酸修饰。单线态氧的寿命足够长(水中有4.4μs)使其能够充分扩散进入细胞和组织。实验室中通常用三种之一的方法来生成单线态氧:1)使用感光染料对二氧(3O2)进行光化学反应来制备;2)通过对一种过氧化物或二氧环丁烷进行加热分解制备;3)氧气流内通过微波放电来制备;单线态氧通过在1270nm处具弱化学发光的典型特征直接被检测。

     

    Thermo fisher(molecular probe)公司开发的Singlet Oxygen Sensor Green®, SOSG是目前商业化可得且应用最广泛的单线态氧荧光探针,从材料学到药物学领域,比如光动力学疗法。

     

    【特异性强】

    Singlet Oxygen Sensor Green®, SOSG高度选择性结合单线态氧(1O2)。不像其他的荧光和化学发光的单线态氧检测试剂,SOSG与其他活性氧物质(ROS),包括羟基自由基(·OH)、超氧阴离子(·O2–)和一氧化氮(NO)无可见反应【见图1】。

    图1、Singlet Oxygen Sensor Green (SOSG)的荧光反应和对单线态氧(1O2)的特异性。A)用荧光光度计(Ex/Em= 488/525nm)测定溶液内的荧光,溶液内含SOSG(1 μM )和亚甲基蓝(10 μM)溶于100mM pH 7.5 Tris Buffer; 单线态氧清除剂叠氮钠(NaN3,1 mM);或50% D2O(重水,能够延长1O2的寿命)。每20s完成一次荧光测定,每次测定间隔30s(如灰条标示)。在这30s的间隔中,样本暴露于激光辐射(630–680 nm,<5 mW),导致亚甲基蓝光敏生成1O2。B)用荧光光度计(Ex/Em=488/525nm)测定溶液内的荧光,溶液为50mM pH 7 Tris buffer,1mM 黄嘌呤,SOSG(1 μM ),或含SOSG(1 μM ),或含二氢罗丹明123。约20s之后,加入50 mU/mL黄嘌呤氧化酶(XO)。XO催化黄嘌呤的氧化,产生尿酸和超氧化物。超氧化物瞬间被H2O2降解。

     

    【荧光特征】

    Singlet Oxygen Sensor Green®, SOSG与单线态氧(1O2)反应前,探针最初发弱蓝色荧光,激发峰在372nm和393nm,发射峰在395nm和416nm。当含单线态氧(1O2),探针发射绿色荧光,光谱特征类似于荧光素(最大激发/发射波长约504/525nm)。

     

    【线性化关系】

    研究发现Singlet Oxygen Sensor Green®, SOSG的荧光产物在某些溶液中随着时间被降解,且在不含单线态氧的碱性pH溶液中,Singlet Oxygen Sensor Green®, SOSG会发荧光。不过,当进行适当的操作,绿色荧光信号的强度与单线态氧(1O2)含量相关,且不会受其他ROS的明显干扰。

     

    【应用】

    Singlet Oxygen Sensor Green®, SOSG证实能用来检测溶液和植物组织中的单线态氧(1O2)。

     

    【使用方法】

    1.       SOSG储存液制备:收到货按照≤-20℃避光干燥保存。用甲醇来制备储存液,每100μg粉末加入33μl甲醇制备~5mM储存液。正式实验前制备工作液,不能用完的工作液需丢弃,不可保存后再用。

    2.       溶液检测:SOSG不具细胞膜渗透性,设计用来检测溶液环境内的单线态氧。最佳的稀释缓冲液和工作浓度需根据经验来调整。建议起始工作浓度为1-10μM。

     

    【使用方法-来自文献】

    一、文献来源: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.

    图2. 肌红蛋白加入脂质体后随着时间的SOSG荧光变化(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.

    图3. 拟南芥叶子内SOSG荧光产生的动力学变化(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、文献来源: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.

    图4. PEG处理后单线态氧和碘化丙啶细胞渗透性的变化图(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.

     

    【争议点】

    Singlet Oxygen Sensor Green®, SOSG不具有细胞膜渗透性,能用于细胞单线态氧(1O2)检测?

    专利持有方thermo(molecular probe)官方说明此探针不具有细胞膜渗透性,适用于溶液体系单线态氧(1O2)检测。然而,Gollmer A et al.报道SOSG能插入活哺乳动物细胞(文献来源: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.)。

     

    订购信息:Thermo fisher原装正品,大包装拆开单卖,100μg /支,单支特价700元。懋康生物长期现货供应,欢迎咨询,QQ:2971634497,或电话:021-54736159。

    品牌

    货号

    名称

    规格

    特价(元)

    Thermo fisher

    Invitrogen

    S36002

    Singlet Oxygen Sensor Green 单线态氧荧光探针

    100μg

    700

    单线态氧研究关联产品

    Maokangbio

    MX4821-1G

    1,3-diphenylisobenzofuran (DPBF) 1,3-二苯基异苯并呋喃(单线态氧指示剂)

    1g

    320

    Maokangbio

    MX4822-50MG

    9,10-Anthracenediyl-bis(methylene)dimalonic Acid (ABDA)  9,10-蒽二基-双(亚甲基)二丙二酸(单线态氧指示剂)

    50mg

    990

    Maokangbio

    MS4075B-1G

    Rose Bengal,95% 孟加拉玫瑰红

    1g

    382

    Maokangbio

    MS5557-50MG

    Chlorin e6 (Ce6)  二氢卟吩e6

    50mg

    972


    【温馨提示】:见我司懋康生物整理的活性氧检测探针专题(Probes for reactive oxygen species)。

     

    相关产品:ROS研究常用的配套产品,需要了解更多信息欢迎来电咨询,021-54736159,QQ:2971634497。

    货号

    名称

    规格             

    价格(元)  

    MX4801-1KIT

    Reactive Oxygen Species (ROS) Assay Kit

    活性氧(ROS)检测试剂盒

    100~500T

    328

    MX4804-1MG

    Aminophenyl fluorescein (APF) 氨基苯基荧光素

    1mg

    2180

    MX4805-1MG

    Hydroxyphenyl fluorescein (HPF) 羟苯基荧光素

    1mg

    2180

    MX4802-50MG

    H2DCFDA (DCFH-DA, DCFH) 活性氧(ROS)荧光探针           

    50mg

    546

    M36008(Invitrogen)    

    MitoSOX Red Mitochondrial Superoxide Indicator

    50 μg

    650

     

    注意事项

    为了您的安全和健康,请穿实验服并戴一次性手套操作。

     

     

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

     

     

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