荧光法胞内pH检测试剂盒

细胞内pH变化涉及多种生理和病理过程，包括细胞增殖，细胞凋亡，受精，恶性肿瘤，多药耐药性，离子转运，溶酶体贮积症和阿尔茨海默病。

Cell Meter™荧光法胞内pH检测剂盒(Cell Meter™ Fluorimetric Intracellular pH Assay Kit)利用专有的荧光指示剂测量相对细胞内pH变化。

Cell Meter™荧光法胞内pH检测剂盒是一种均相的，动力学的活细胞荧光测定法，它使用标准程序或酸负荷程序。设计标准方案用于治疗靶标，同时治疗时细胞内pH降低。“酸负荷”程序旨在测量与GPCR活化或生长因子活性引起的细胞代谢变化相关的细胞内pH的增加。在“酸负荷”程序中，在将荧光pH染料以最小体积加载到细胞中之后加入氯化铵溶液。

该“酸加载”步骤之后是在相对大体积（~4X）的缓冲液中加入激动剂。突然的体积变化引发来自细胞的氨（NH 3）流出，导致细胞内pH的快速降低，并因此导致荧光信号的降低。通过相对荧光信号增加测量激动剂对随后的细胞内pH恢复的影响。

Intracellular pH change are implicated in diverse physiological and pathological processes, including cell proliferation, apoptosis, fertilization, malignancy, multidrug resistance, ion transport, lysosomal storage disorders and Alzheimer's disease. The Cell Meter™ Fluorimetric Intracellular pH Assay Kit utilizes AAT Bioquest's proprietary fluorescent indicator for measuring the relative intracellular pH changes. It is a homogeneous, kinetic, live-cell fluorescent assay that utilizes either a standard procedure or acid-load procedure. The standard protocol is designed for measuring the therapeutic targets of interest with a decrease in intracellular pH upon treatment. The 'Acid-Load' procedure is designed to measure the increase of intracellular pH associated with changes in cellular metabolism due to GPCR activation or growth factor activity. With the 'Acid-Load' procedure ammonium chloride solution is added after the fluorescent pH dye is loaded into cells in a minimum volume. This 'acid-loading' step is followed by the addition of agonist in a relatively large volume (~4X) of buffer. The sudden volume change initiates an efflux of ammonia (NH3) from the cells causing a rapid decrease in intracellular pH, and thus a decrease in fluorescence signal. The effect of agonist on the subsequent recovery of intracellular pH is measured by the relative fluorescence signal increase.

适用仪器

>>荧光酶标仪  

Ex: 490 nm

Em： 535 nm

Cutoff： 515 nm

推荐孔板: 黑色透明底板

>>其他可用仪器  

FDSS, FLIPR, ViewLux, NOVOStar, ArrayScan, FlexStation, IN Cell Analyzer