5(6)-羧基SNARF-1,乙酰甲酯, 乙酸酯

产品描述

透过细胞的pH指示剂，羧基SNARF-1，乙酰氧基甲酯，乙酸酯（CAS＃126208-13-7）在脱酯化后的pKa为7.5，因此可用于测量pH在7和pH 8之间的变化。羧基-SNARF-1在酸性和碱性条件下分别显示出从黄橙色到深红色荧光的明显的pH依赖性发射变化。这种对pH的依赖性使得染料在两个发射波长（通常为580 nm和640 nm）处的荧光强度之比可用于定量测定pH。避光保存于-20℃。

用SNARF AM醋酸酯加载细胞

研究人员应确定每种细胞类型和实验的最佳加载条件。文献援引了各种各样的上样条件，从1到20 µM SNARF AM醋酸盐与细胞一起温育10到60分钟。 DMSO储备溶液通常至少以1：1000的比例稀释到上样缓冲液中，以减少细胞对DMSO的暴露，并且上样缓冲液应无血清，因为血清通常含有酯酶活性。非离子型洗涤剂Pluronic F-127有时用于促进非极性SNARF AM乙酸酯分散到缓冲液中。

作为初始上样条件，我们建议将细胞在1~10 µM SNARF AM醋酸盐中孵育30分钟，并在最合适的温度下孵育特定的目标细胞类型。上样后，应在开始pH测量之前清洗细胞。为了装载脑片，建议在含有20µM羧基SNARF-1 AM醋酸盐和4％（w / v）Pluronic F-127的人工脑脊液（ACSF）中孵育60分钟，然后再在无染料ACSF中孵育30分钟。

Product Description

The cell-permeant pH indicator, carboxy SNARF-1, acetoxymethyl ester, acetate (CAS #126208-13-7) has a pKa of ~7.5 after de-esterification, thus is useful for measuring pH changes between pH 7 and pH 8. Carboxy-SNARF-1 exhibits a significant pH-dependent emission shift from yellow-orange to deep red fluorescence under acidic and basic conditions, respectively. This pH dependence allows the ratio of the fluorescence intensities from the dye at two emission wavelengths – typically 580 nm and 640 nm – to be used for quantitative determinations of pH. Store at -20℃ in the dark.

Loading Cells with SNARF AM Acetate Esters

Optimal loading conditions for each cell type and experiment should be determined by the researcher. The literature cites a wide range of loading conditions, from 1 to 20 µM SNARF AM acetate incubated with cells for from 10 to 60 minutes. DMSO stock solutions are typically diluted at least 1:1000 into loading buffer to reduce the exposure of cells to DMSO, and the loading buffer should be se rum-free because serum often contains esterase activity. The non ionic detergent Pluronic F-127 is some times used to promote dispersion of the rather nonpolar SNARF AM acetate esters into buffers. 

As initial loading conditions, we recommend incubating cells in 1～10 µM SNARF AM acetate for 30 minutes at the optimum temperature for the specific cell type of interest. After loading, cells should be washed before commencing pH measurements. For loading brain slices, incubation for 60 minutes in artificial cerebrospinal fluid(ACSF) containing 20µM carboxy SNARF-1 AM acetate and 4% (w/v) Pluronic F-127 followed by a further 30 minute incubation in dye-free ACSF is recommended.