| 包装: | 1mg |
| 市场价: | 2374元 |
C11-BODIPY581/591 是一种用于脂质氧化的荧光比率探针。
Cell lines | microalgal cells (unicellular green alga Chlamydomonas reinhardtii); mammalian cells |
Preparation method | The stock solution of C11-BODIPY581/591 was prepared by dissolving 1 mg of the product in DMSO to a concentration of 1mM. Stock solution can be stored in freezer (-5 to -30℃) and protect from light. |
Reaction Conditions | 2.5 μM C11-BODIPY581/591 for 30 min. The excitation and emission band of oxidized type is pass of 460–495 and 510–550, respectively. But the excitation and emission band of reduced type is pass of 565–581 and 585–591, respectively. |
Applications | C11-BODIPY591/581 staining was combined with flow cytometry measurements to detect reactive oxygen species (ROS) and oxidative stress damage in living cells and membrane systems. Oxidation of the polyunsaturated butadienyl portion of the dye results in a shift of the fluorescence emission peak from ~590 nm to ~510 nm. |
Cell lines | Rat-1 fibroblasts |
Preparation Method | Rat-1 fibroblasts were cultured at 37℃ in DMEM supplemented with 7.5% fetal calf serum in a 7.5% CO2humidified atmosphere. Small unilamellar vesicles of egg PC were made through ethanolic injection. The C11-BODIPY581/591concentration of the stock solutions was determined by measuring the absorption at 581 nm using a molar extinction coefficient of 139 444 l mol-1cm-1. |
Reaction Conditions | Cells were incubated for 30 min at 37℃ with C11-BODIPY581/591(1μM) in growth medium. Prior to use, fibroblasts were rinsed with enriched phosphate buffered saline (PBS+, 137 mM NaCl, 2.7 mM KCl, 0.5 mM MgCl2, 0.9 mM CaCl2, 1.5 mM KH2PO4, 8.1 mM Na2HPO4and 5 mM glucose, pH 7.4). Oxidation of C11-BODIPY581/591was induced by incubating cells with 50μM CumOOH at 37℃ with or without hemin in PBS+. |
Applications | C11-BODIPY581/591is oxidized only when CumOOH and hemin are present. The oxidation rate correlates directly with CumOOH concentration, up to 0.2 mM. Administration of C11-BODIPY581/591dissolved in fetal calf serum to the growth medium of rat-1 fibroblasts leads to a rapid incorporation into cellular membranes. When the ratio of C11-BODIPY581/591to cellular phospholipid is 1:13 047±734, it causes the minimal perturbation of the membranes and the absence of spectroscopic artifacts after 30 min of labeling. |
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| 产品描述 | C11-BODIPY581/591is an oxidation-sensitive fluorescent fatty acid analogue with fluorescent properties in the red range of the visible spectrum (emission maximum 595 nm), allowing its application in fluorescence microscopy. C11-BODIPY581/591is easily incorporating into membranes and fluoresces red in the intact state but shifts to green upon free radical-induced oxidation. This characteristic is highly advantageous, it makes the ratio-imaging of oxidant activities at the (sub)cellular level feasible. In addition, the fluorescent properties of C11-BODIPY581/591allow the use of this probe in fast- and medium- throughput screening of antioxidants in living cells and model membranes in a multiwell/fluorescence reader approach[1][2]. The wavelengths of maximal excitation and emission of fluorophore C11-BODIPY581/591corresponded to 581 and 591 nm, respectively. Addition of CumOOH/hemin, as an initiator of lipid oxidation, shifted the excitation and emission spectra to shorter wavelengths corresponding to green fluorescence (peak excitation 500 nm, emission 510 nm). C11-BODIPY581/591is also easily oxidized by other hydroxy-, peroxy- and oxy-radical generating systems such as hydrogen peroxide/Fe2+ and 2,2’-azobis. However, this probe is relatively insensitive to SIN-1, which generates nitric oxide and superoxide[3] References: |
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