Fluo3是一种长波钙荧光探针.doc

Product Name: Fluo 3-AM Chemical Name: 1-[2-Amino-5-(2,7-dichloro-6-hydroxy-3-oxo-9-xanthenyl)phenoxy]-2-(2-amino-5- methylphenoxy)ethane-N,N,N,N-tetraacetic acid, pentaacetoxymethyl ester CAS Number: 121714-22-5 C51H50Cl2N2O23 = 1129.85 Product Description: Storage: -20 oC Shipping Condition: Ambient Temperature Appearance: red solid Fluorescent spectrum: pass test Solubility: 2 mg/ml acetonitrile, 0.9 mg/ml DMSO Fluo3是一种长波钙荧光探针。Fluo3若以游离配体形式存在时几乎是非荧光性的，但是当它与钙离子结合后荧光会增加60至80倍。因为共聚焦激光荧光显微镜具有氩激光器，所以Fluo3被广泛使用于这种显微镜上。这种荧光信号发出来的长波也便于减小对样品细胞的光损伤。Fluo3对于紫外光照射下可裂解的螯合钙或其它形式的钙也有作用。Fluo 3-AM是Fluo3的一种乙酸甲酯衍生物，很容易通过孵育导入细胞中。 Protocol: 1. Add 16.5 mg Pluronic F127 to Fluo 3-AM/DMSO solution. Pluronic F127 prevents aggregation of Fluo 3-AM in HBSS and helps uptake with cells. 2. Dilute the Fluo 3-AM solution with HBSS to prepare 4 mM Fluo 3-AM working solution. 3. Add the Fluo 3-AM working solution to the cells, and incubate at 37 oC for 20 min. 4. Add 5 volumes of HBSS containing 1% fetal calf serum, and continue the incubation for another 40 min. 5. Wash the cells 3 times with HEPES buffer saline. Then resuspend the cells to prepare 1x105 cells/ml solution using HEPES buffer saline. 6. Incubate at 37 oC for 10 min. Then use the cells for fluorescent calcium ion detection. 7. Monitor the fluorescence at 528 nm (excitation: 490-500 nm). *Cell staining conditions differ by cell types, so it is necessary to optimize the conditions for each experiment. DOJINDO LABORATORIES 同仁化学研究所 中国（上海）代表处地址: 上海市零陵路629号爱和大厦25楼H座电话:021 021URL：E-mail:info@