常用荧光探针小结.pptVIP

常用荧光探针小结 一、异硫氰酸荧光素（Fluorescein isothiocyanate, FITC） FITC有两种异构体，性质稳定，低温下干燥保存，其性状多年不变，室温下也能保存两年以上。异构体I、II均能与蛋白质良好结合，但异构体I的荧光效率更高，与蛋白质的结合也更稳定。 FITC的最大吸收光谱为490----495纳米，最大发射光谱为520-530nm，呈明亮的黄绿色荧光。FITC含有异硫氰基，在碱性条件下能与IgG的自由氨基（主要是赖氨酸的?-氨基）形成荧光抗体结合物。 * Exitation λmax: 495 nm; Emission λmax: 519 nm; Solvent pH:8.00 * * Immunocytochemistry/Immunofluorescence-alpha Tubulin antibody [DM1A] (FITC) (ab64503) ICC/IF image of ab64503 stained human HeLa cells. The cells were methanol fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab64503, 1μg/ml, FITC conjugated (green)) for 1h at room temperature. 1% BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. Alexa Fluor? 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). * 二、荧光素酯 Carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) is a cell permeable dye generally used in animal cell proliferation research. CFDA-SE enters cells by diffusion and is cleaved by intracellular esterase enzymes to form an amine-reactive product, CFSE. This product produces a detectable fluorescence and covalently binds to intracellular lysine residues and other amine sources. 酯酶 * 视频：The Use of Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) to Monitor Lymphocyte Proliferation * 四甲基异硫氰酸罗达明，它是一种紫红色粉末，较稳定。其最大吸收光谱为550nm，最大发射光谱620nm，呈橙红色荧光，与FITC的黄绿色荧光对比清晰，与蛋白质结合方式同TITC。它可用于双标记示踪研究。 三、异硫氰酸罗丹明（TMRITC） * Detection of α-tubulin in A549 cells demonstrates use of rhodamine-labeled secondary antibody. Cells were probed with a mouse anti-α-tubulin primary antibody (0.4μg/mL) and Rhodamine-goat anti-mouse secondary antibody (2μg/mL). Nuclei were labeled with Hoechst Dye. Images were acquired by fluorescence microscopy. A. Fluorescence image shows a delicate network of α-tubulin (pseudo-colored green) located exclusively in the cytoplasm. B. Nuclear counterstain with Hoechst Dye (pseudo-colored blue) C.