JC-1线粒体膜电位荧光探针

¥3,000.00

规格: 5mg

库存详情未设置 SKU: MB6055-1 分类: , ,
JC-1线粒体膜电位荧光探针
JC-1在通过流式细胞术检测线粒体膜电位中广泛使用。它能够选择性的进入线粒体中,并且由于膜电位的增加(大约超过80-100 mV)其颜色会发生从绿色到橙色的可逆变化。这种特性是由于JC-1聚合物的可逆结构,在膜极化条件下,它的发射光由530nm(J-单体发射光)转移到590nm (JC-1聚合体发射光)。当在490nm处激发时,JC-1发生从绿色到橙绿色的可逆改变,因为线粒体膜极化的更大。这两种颜色都可以被流式细胞仪上装好的一般滤光器检测到,因此绿色发射光可以通过荧光通道1(FL1)进行分析,橙绿色发射光可以通过荧光通道2(FL2)进行分析。JC-1主要的优点是,它既可以进行定性检测,通过其发射荧光的颜色从绿色到橙色的改变;也可以进行定量检测,通过对两个检测通道FL1和FL2单一色彩的荧光强度进行检测;它也可以用于荧光成像分析。We have developed a protocol to use it in fluorescence microplate platform.。虽然JC-1广泛地用于许多实验中,但是其水溶性差,也导致它很难在一些实验中使用。我们的JC-10的水溶性比JC-1要高,并且在许多细胞系中有着超好的性能表现;有趣地是,JC-10的性能表现对细胞系极具依赖性。
规格
5 mg
产品形式
粉末
Ex (nm)
515
Em (nm)
529
分子量
652.23
溶剂
DMSO
储存条件:-20℃
操作说明(仅供参考)
1. Prepare JC-1 working solution:
1.1 Prepare a 2 to 10 mM stock solution of JC-1 in high-quality, anhydrous DMSO. The stock solution should be used promptly; any remaining solution should be aliquoted and frozen at<-20oC.
Note: Avoid repeated freeze-thaw cycles, and protect from light.
1.2Prepare a 1X JC-1 working solution: On the day of the experiment, either dissolve JC-1 solid in DMSO or thaw an aliquot of the JC-1 stock solution to room temperature. Prepare a 10 to 30 µM 1X JC-1 working solution in Hanks and 20 mM Hepes buffer (HHBS) or buffer of your choice, pH 7 with 0.02% Pluronic® F-127(MA0061). Mix them well by votexing.
Note: JC-1is not water soluble, so it intends to aggregate in solution. It is recommended to filter the JC-1 working solution before loading it into the cells.
2. Run JC-1 assay with a fluorescence microplate reader:
2.1Treat cells with test compounds for a desired period of time (For example, Jurkat cells can be treated with camptothecin for 4-6 hours) to induce apoptosis. For blank wells (medium without the cells), add the corresponding amount of compound buffer.
2.2Add 100 µL/well/96-well plate or 25 µL/well/384-well plate of JC-1 working solution (from Step 1.2) into the cell plate.
2.3 Incubate the JC-1 loading plate in a 37oC, 5% CO2incubator for 15-60 min.
Note: The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment.
2.4Remove the JC-1 working solution from the plate, wash the cells with HHBS or buffer of your choice. Add 100 µL/well/96-well plate or 25 µL/well/384-well plate of HHBS back to the cell plate.
2.5Monitor the fluorescence change atEx/Em = 490/525 nm and 490/590 nm for ratio analysis.
3. Run JC-1 assay with a fluorescence microscope or a flow cytometer:
3.1Treat cells with test compounds for a desired period of time (For example, Jurkat cells can be treated with camptothecin for 4-6 hours) to induce apoptosis.
3.2Centrifuge the cells to get 1-5 x 105cells per tube.
3.3 Resuspend cells in 500 µL of JC-1 working solution (from Step 1.2).
3.4Incubate at room temperature or 37° C for 10 to 30 min, protected from light.
3.5Wash the cells with HHBS or buffer of your choice. Resuspend cells in 500 µL of HHBS to get 1-5 x 105cells per tube.
3.6Monitor the fluorescence change at Ex/Em = 490/525 nm and 490/590 nm with a fluorescence microscope (using FITC and TRITC filters) or a flow cytometer (using FL1 and FL2 channels).
相关产品推荐
MB6097
JC-10线粒体膜电位荧光探针
MA0061
Pluronic™ F-127 (20% Solution in DMSO)
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规格

5mg

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