Page 7 - 2016 July 專刊-H-0722

偵測粒線體膜電位改變

偵測粒線體膜電位
【Mitochondrial Membrane Potential Kit】SI-MAK159 / SI-MAK160

偵測原理
細胞凋亡初期，線粒體轉膜電位會發生變化， 實驗操作流程
導致膜穿透性的改變。利用是陽離子染劑JC-
10，可以偵測到細胞膜的通透改變。JC-10在
正常細胞中，會在線粒體中形成聚集體，發出
強烈的紅色螢光。但是在凋亡的細胞中，因線
粒體穿膜電位的改變，會以單體形式存在細胞
液中，而發出綠色螢光。

Add 50ul of JC-10 dye ,
and incubate 30~60 min.
Then add 50 ul of assay
buffer B
Monitor the fluorescence
intensity (lex = 490/
lem = 525 nm) and (lex =
540/lem = 590 nm) for
ratio analysis. The ratio
of red/green fluorescence
intensity is used to
determine MMP.

Campotothecin-induced mitochondria membrane potential Effect of FCCP induced mitochondria membrane
changes were measured with JC-10 and JC-1 in Jurkat potential change in JurKat cells. JurKat cells were
cells. After Jurkat cells were treated with camptothecin dye loaded with JC-10 dye-loading solution along
(10 μM) for 4 hours, JC-1 and JC-10 dye loading with DMSO (Top) or 5 μM FCCP (Low) for 10
solutions were added to the wells and incubated for 30 minutes. The fluorescent intensities for both J-
minutes. The fluorescent intensities for both J-aggregates aggregates and monomeric forms of JC-10 were
and monomeric forms of JC-1 and JC-10 were measured measured with a flow cytometer using FL1 and FL2
at Ex/Em = 490/525 nm and 490/590 nm. channels. Uncompensated data (left column) were
compared with compensated data (right column).

相 關 產 品 偵 測 方 式 產 品 品 號 包 裝

Mitochondria Membrane Potential Microplate Readers SI-MAK159 500Test
Kit (JC-10 assay) Flow SI-MAK160 100Test

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