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1392488-04-8
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NIR-H2O2
CAS No. : 1392488-04-8
MCE 国际站：NIR-H2O2
产品活性：NIR-H2O2 是一种可透过细胞的 NIR 荧光启动传感器。NIR-H2O2 在 NIR 区域内同时具有吸收和发射作用。当在 NIR 区域激发时，NIR-H2O2 对 H2O2 产生较大的启动 NIR 荧光信号。NIR-H2O2 能够对活细胞和小鼠中内源性产生的 H2O2 进行成像。
研究领域：Others
作用靶点：Fluorescent Dye
In Vitro: NIR-H2O2 is highly selective to H2O2 over other typical ROS and biorelevant species.
HeLa cells incubated with NIR-H2O2 (5 μM) for 30 min at 37 °C provide almost no fluorescence. However, when the living HeLa cells loaded with NIR-H2O2 are further treated with H2O2, they give strong fluorescence. NIR-H2O2 is cell membrane permeable and responsive to H2O2 in the living cells. When stimulated by phorbol myristate acetate (PMA), macrophage cells may produce endogenous H2O2. The living RAW264.7 macrophage cells loaded with only the NIR sensor NIR-H2O2 (1 μM) display almost no fluorescence. However, the macrophage cells coincubated with PMA (3.0 μg/mL) and the sensor NIR-H2O2 (1 μM) exhibit a dramatic enhancement in the red emission. NIR-H2O2 is capable of fluorescent imaging of endogenously produced H2O2 in the living RAW264.7 macrophage cells. The mitochondria staining experiments suggest that the sensor mainly associates with the mitochondria of RAW264.7 macrophage cells.
In Vivo: The H2O2 production in vivo was generated by activated macrophages and neutrophils in a lipopolysaccharide (LPS) model of acute inflammation. The mice treated with both LPS and NIR-H2O2 exhibit a significantly higher fluorescence readout than the mice untreated or treated with only NIR-H2O2. The mice loaded with LPS and NIR-H2O2 have approximately 10- and 20-fold higher fluorescence intensity than the mice loaded with saline and the sensor and the mice loaded with saline, respectively.
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文献和实验信号也很重要。在确定哪种显微镜物镜可以分辨标本的最小特征时，就需要考虑数值孔径。在权衡选项利弊时，请记住数值孔径范围在 0.04 至 1.7 之间。 3. 图像视场和景深要求是什么？ 视场数或视场直径是光学显微镜中视场的直径，以毫米为单位，并在中间像平面测量。现代平场复消色差透镜和其他专用平场物镜通常具有 22 到 26.5 毫米（与广角目镜组合使用时或会更大）的可用视场。 物镜的景深即在图像锐度不发生明显变化情况下的物镜轴向聚焦范围。该值从低数值孔径物镜到高数值孔径物镜变化很大；通常随着数
化物，室温10～20分钟。4．正常血清（1:10）孵育，室温20分钟。5．滴加第一抗体，37℃1小时或4℃过夜。6．PBS洗涤3×3分钟。7．滴加酶标二抗，室温1小时。8．PBS洗涤3×3分钟。9．0.04%DAB＋0.03 %H2O2显色5～10分钟。10． 充分水洗后，复染、脱水、透明、封片、镜检。三、PAP法（过氧化物酶抗过氧化物酶抗体复合物法）1．标定固定后，PBS洗涤。2．1% H2O2（或1% H2O2甲醇）阻断内源性过氧化物，室温10～20分钟。3．PBS洗涤2×3分钟。4．正常血清（二抗
去缓冲液A，滴加标记液约50μl，25℃温育1h。标记反应液：0.005mmol/L dNTP, 0.005mol/L biotin-16-dUTP, 25U/ml Klenow大片段。 6．PBS漂洗2次，各3min。 7．内源酶阻断剂阻断15min。 8．PBS洗2次，各3min。 9．滴加HRP-avidin覆盖组织，室温下30min。 10．PBS洗2次，各3min。 11．DAB-H2O2显色约5min。 12．流水冲洗后，苏木素复染1min。常规脱水，透明
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