常用荧光探针小结
25页1、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。,1,Exitation max: 495 nm; Emission max: 519 nm; Solvent pH:8.00,2,3,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
2、% PBS-Tween (20 min) and incubated with the antibody (ab64503, 1g/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).,4,二、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 ce
3、lls 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,5,视频:The Use of Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) to Monitor Lymphocyte Proliferation,6,四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),7,Detection of -tubulin in A549 cel
4、ls demonstrates use of rhodamine-labeled secondary antibody. Cells were probed with a mouse anti-tubulin primary antibody (0.4g/mL) and Rhodamine-goat anti-mouse secondary antibody (2g/mL). Nuclei were labeled with Hoechst Dye. Images were acquired by fluorescence microscopy. A. Fluorescence image shows a delicate network of -tubulin (pseudo-colored green) located exclusively in the cytoplasm. B. Nuclear counterstain with Hoechst Dye (pseudo-colored blue) C. Merged image.,8,AC41323-0010,RB200,也称
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