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線粒體膜電位熒光探針JC-10 JC-1的卓越代替品

貨號22204存儲條件 在零下15度以下保存
規格5x100 uL價格1236
Ex (nm)508Em (nm)524
分子量583.34溶劑DMSO
產品詳細介紹


簡要概述

線粒體膜電位熒光探針JC-10是美國AAT Bioquest研發的用于檢測線粒體膜電位的熒光探針是JC-1的完美替代品。JC-1在許多實驗室中被廣泛使用,但其水溶性差使得它在某些應用中難以使用。即使在1μM濃度下,JC-1也傾向于在水性緩沖液中沉淀。當需要高染料濃度時,JC-10已被開發為JC-1的替代物。與JC-1相比,我們的JC-10具有更好的水溶性。 JC-10能夠選擇性地進入線粒體,并隨著膜電位的增加可逆地將其顏色從綠色變為橙色。這種性質是由于膜極化時JC-10聚集體的可逆形成導致發射光從520nm(即JC-10單體形式的發射)轉變為570nm(即J-聚集體形式的發射)。當在490nm激發時,隨著線粒體膜變得更加極化,JC-10的顏色從綠色橙色可逆地變為綠色橙色。使用通常安裝在所有流式細胞儀中的過濾器可以檢測兩種顏色??梢栽跓晒馔ǖ?(FL1)中分析綠色發射,在通道2(FL2)中分析綠色橙色發射。除了用于流式細胞儀外,JC-10還可用于熒光成像。我們已經開發出一種在熒光微孔板平臺中使用JC-10的方案。在一些細胞系中,JC-10具有優于JC-1的性能。百螢生物是AAT Bioquest 的中國代理商,為您提供最優質的線粒體膜電位熒光探針。

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產品說明書

JC-10的分析方案

概述

準備含有測試化合物的細胞

添加JC-10工作溶液(100μL/孔用于96孔板或25μL/孔用于384孔板)

在室溫或37 ℃孵育1小時

在Ex讀取熒光強度 / Em = 490 / 525nm和540 / 590nm

注意:以下是我們推薦的活細胞方案。 該協議僅提供指南,應根據您的特定需求進行修改。

 

操作步驟

1.準備JC-10工作溶液:

1.1每瓶DMSO原液(100μL,2 mg / mL,3 mM)只能使用一次。任何未使用的小瓶應儲存在<-20℃。注意:避免反復凍融循環,并避光.

1.2準備1X JC-10工作溶液:在實驗當天,解凍一份JC-10 stockolution到室溫。在Hanks和20 mM Hepesbuffer(HHBS)或您選擇的緩沖液(pH 7-8,含0.02%Pluronic [表情] F-127)中制備10至30μM1X工作溶液。通過votexing將它們混合均勻。注意:對于某些細胞系,pH值為8的工作溶液可能會阻止JC-10泄漏。

2.用熒光酶標儀進行JC-10檢測:

2.1用測試化合物處理細胞一段所需的時間(例如,Jurkat細胞可以用喜樹堿處理4-6小時)以誘導細胞凋亡。 對于空白孔(沒有細胞的培養基),加入相應量的化合物緩沖液。

2.2將100μL/孔/ 96孔板或25μL/孔/ 384孔板的JC-10工作溶液(來自步驟1.2)加入到細胞板中。

2.3將JC-10加載板在37 oC,5%CO2培養箱中孵育15-60分鐘。

注意:適當的孵育時間取決于所使用的單個細胞類型和細胞濃度。優化每個實驗的孵育時間。

2.4監測Ex / Em = 490/525 nm(FITC通道)和540/590 nm(TRITC通道)的熒光變化,進行比率分析。

可選:從板上取下JC-10工作溶液; 在分析之前,將100μL/孔/ 96孔板或25μL/孔/ 384孔HHBS板加回到細胞板中。

3.用熒光顯微鏡或流式細胞儀進行JC-10檢測:

3.1用測試化合物處理細胞一段所需的時間(例如,Jurkat細胞可以用喜樹堿處理4-6小時)以誘導細胞凋亡。

3.2離心細胞,每管取1-5×105個細胞。

3.3將細胞重懸于500μLJC-10工作溶液中(來自步驟1.2)。

3.4在室溫或37°C,5%CO2培養箱中孵育10至30分鐘,避光。

注意:適當的孵育時間取決于所使用的單個細胞類型和細胞濃度。優化每個實驗的孵育時間。

3.5使用熒光顯微鏡(使用FITC和TRITC過濾器)或流式細胞儀(使用FL1和FL2通道)監測Ex / Em = 490/525 nm和540/590 nm處的熒光變化??蛇x:移除JC-10工作 從板上解決; 在熒光顯微鏡下分析之前,將100μL/孔/ 96孔板或25μL/孔/ 384孔HHBS板加回到細胞板中。

 

參考文獻

JC-10™ has been used to study many biologically significant processes across several key disciplines. To list a few, JC-10™ has been used to investigate topics such as mitochondrial membrane potential, cytotoxicity, cell viability, oxidative stress, cancer metastasis, apoptosis, signal transduction, mitochondrial fission and induced pluripotent stem cells (iPSCs). 

Below, you may find a small sampling of specific JC-10™ applications. To inquire about a potential application of JC-10™, or to consult with our fluorescent dye specialists, please contact us at support@aatbio.com or 1-800-990-8053.

High-content assays for hepatotoxicity using induced pluripotent stem cell–derived cells.
Researchers use JC-10™ to monitor mitochondrial depolarization as an indication of hepatotoxicity and oxidative stress in induced pluripotent stem cell-derived cells, with the ultimate goal of designing a reliable, high-content and imaging-based in vitro toxicity assay. 

High-Content High-Throughput Assays for Characterizing the Viability and Morphology of Human iPSC-Derived Neuronal Cultures
JC-10™ was used to study neurons derived from induced pluripotent stem-cells. Since JC-10™ will accumulate in the mitochondria of viable cells, it was used to determine cell viability in a high-throughput assay context.

Anticancer Activity of New Synthetic α-Methylene-δ-Lactones on Two Breast Cancer Cell Lines
Researchers chose JC-10™ to investigate mitochondrial membrane potential and membrane integrity in cells treated with natural products, such as α-Methylene-δ-Lactones, with the goal of developing new treatments for breast cancer.

Tetrandrine protects mouse retinal ganglion cells from ischemic injury.
JC-10™ was used in flow cytometry to study mitochondrial membrane potential (ΔΨm) in primary cultured retinal ganglion cells, as an extension of the field of drug discovery into prevention of ischemic injury. 

Midazolam induces cellular apoptosis in human cancer cells and inhibits tumor growth in xenograft mice
In a study of human cancer cells, JC-10™ was employed to track cellular apoptosis as a function of mitochondrial membrane potential, and consequently, mitochondrial activity. Researchers were interested in the possible anesthetic properties of midazolam for anticancer drug delivery.

Mitochondrial proteomics with siRNA knockdown to reveal ACAT1 and MDH2 in the development of doxorubicin-resistant uterine cancer
JC-10™ was used by researchers for the purposes of drug discovery. In particular, researchers were interested in finding new treatments for doxorubicin-resistant uterine cancer, using JC-10™ to monitor mitochondrial membrane potential and validate cell viability results. 

Cold exposure lowers energy expenditure at the cellular level
Researchers used JC-10™ to investigate the relationship between temperature and cellular activity. In particular, researchers wanted to explore if cold temperature acts as a stressor on mitochondrial membrane potential, with regards to the oxidative phosphorylation process which generates ATP.

Susceptibility of gametes and embryos of the eastern oyster, Crassostrea virginica, to Karenia brevis and its toxins
JC-10™ was used in the study of sperm viability, fertilization successs and embryonic survival of Crassostrea virginica. Specifically, JC-10™ was used to quantify mitochondrial membrane potential in sperm cells and to determine possible toxicity effects of algal blooms.

Calmodulin antagonists induce cell cycle arrest and apoptosis in vitro and inhibit tumor growth in vivo in human multiple myeloma
JC-10™ was used by researchers to study cell cycle and apoptosis in human multiple myeloma. JC-10™ functioned as a probe for the detection of mitochondrial membrane potential depolarization, which was crucial to the study of caspase activated apoptosis.

Activation of the mitochondrial apoptotic pathway produces reactive oxygen species and oxidative damage in hepatocytes that contribute to liver tumorigenesis
Researchers were interested in the pathways involved with liver tumorigenesis. To that end, they used JC-10™ to study activation of apoptotic pathways related to changes in mitochondrial membrane potential, with the ultimate goal of discovering if antioxidant therapy might help suppress liver carinogenesis.



說明書
線粒體膜電位熒光探針JC-10 JC-1的卓越代替品.pdf


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