JC-10 Mitochondrial膜Potential Assay试剂盒(Microplate) (ab112134)

概述

  • 产品名称JC-10 Mitochondrial膜Potential Assay试剂盒(Microplate)
    参阅全部 Mitochondrial Transmembrane Potential 试剂盒
  • 检测方法Fluorescent
  • 测试
    5 x 96 well plate
  • 样品类型
    Adherent cells, Suspension cells
  • 检测类型Direct
  • 产品概述

    Jc-10 Mitochondrial Membrane Potential Assay Kit (Microplate) (ab112134) enables researchers to run JC-10 assay in the format of microplate reader, and it provides the most robust assay method for monitoring mitochondria membrane potential changes. Thsi assay is based on the detection of the mitochondrial membrane potential changes in cells by the cationic, lipophilic JC-10 dye. In normal cells, JC-10 concentrates in the mitochondrial matrix where it forms red fluorescent aggregates. However, in apoptotic and necrotic cells, JC-10 diffuses out of mitochondria. It changes to monomeric form and stains cells in green fluorescence.

    Although JC-1 is widely used in many labs, its poor water solubility causes great inconvenience. Even at 1 µM concentration, JC-1 tends to precipitate in aqueous buffer. JC-10 is developed to be a superior alternative to JC-1 when high dye concentration is desired. Compared to JC-1, JC-10 has much better water solubility. JC-10 is capable of selectively entering into mitochondria, and reversibly changes its color from green to orange as membrane potentials increase. This property is due to the reversible formation of JC-10 aggregates upon membrane polarization that causes shifts in emitted light from 520 nm (i.e., emission of JC-10 monomeric form) to 570 nm (i.e., emission of J-aggregate form). When excited at 490 nm, the color of JC-10 changes reversibly from green to greenish orange as the mitochondrial membrane becomes more polarized. Both colors can be detected using the filters commonly mounted in all flow cytometers. The green emission can be analyzed in fluorescence channel 1 (FL1) and greenish orange emission in channel 2 (FL2). Besides its use in flow cytometry, it can also be used in fluorescence imaging and fluorescence microplate platform.

     

    Visit our FAQs page for tips and troubleshooting.

  • 说明

    A microplate reader with bottom-reading mode is essential to perform this assay.

    If you would like to use JC-10 on a flow cytometer, we recommend JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry) (ab112133).

  • 经测试应用适用于: Functional Studiesmore details
  • 平台Microplate

性能

  • 相关性Mitochondrial Transmembrane Potential is an important parameter of mitochondrial function used as an indicator of cell death. The collapse of the mitochondrial transmembrane potential coincides with the opening of the mitochondrial permeability transition pores, leading to the release of cytochrome c into the cytosol, which in turn triggers other downstream events in the apoptotic cascade.
  • 别名
    • mitochondrial membrane potential

应用

Our Abpromise guarantee covers the use of ab112134 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Functional Studies Use at an assay dependent concentration.

JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate) 图像

  • Camptothecin-induced mitochondria membrane potential changes with JC-10 and JC-1 in Jurkat cells. JC-1 (blue bar) and JC-10 (red bar) dye loading solutions were added to Jurket cells untreated (0 µM) or treated with camptothecin (10 µM for 4 hours) and incubated for 30 minutes. The fluorescent intensities for both J-aggregates and monomeric forms of JC-1 and JC-10 were measured at Ex/Em = 490/525 nm and 490/590 nm with a microplate reader.

实验方案

JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate) (ab112134)参考文献

This product has been referenced in:
  • Jayathilake AG  et al. Krill oil extract suppresses cell growth and induces apoptosis of human colorectal cancer cells. BMC Complement Altern Med 16:328 (2016). Read more (PubMed: 27576529) »
  • Kysenius K  et al. Mitochondria and NMDA receptor-dependent toxicity of berberine sensitizes neurons to glutamate and rotenone injury. PLoS One 9:e107129 (2014). Read more (PubMed: 25192195) »

See all 3 Publications for this product

Product Wall

Thank you for your enquiry.

JC-1, a lipophiliccationic dye, is widely used to detect mitochondrial de-polarization. However it has poor water solubility which means it precipitates in aqueous buffer even at 1 μM concentration. Compared to J...

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You will not remove the treatment media prior to step C3.
The final volume in the well is 200 uL (100 uL cells + 50 uL JC-10 + 50 uL assay buffer B).

Assay Buffer B is used to quench the reaction and reduce the overall background. It is not necessary to perform the assay successfully however, it will definitely improve the results.

Phenol red should be OK; in our experience there has not been any interference.

The concentration of 100X JC-10 in ab112134 is 3 mM. The solubility is ˜ 5mg/mL in DMSO.



I can confirm that the numbers seems to be alright to me.

The product is used for mitochondria membrane potential measurement.

The dye needs to get into the live cells to exhibit either green (in cytosol) or red (in mitoch...

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Thank you for contacting us about ab112134. Here is some further information: - The blank contains no cells and is growth media only. This gives the background and should be subtracted from all readings. - The control sample should have cells wi...

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Yes, you can do it, although the background might increase a bit.

Thank you for your message.

I understand the confusion and apologize for the inconvenience.

We have checked this and the documents have been amended with the correct information: We recommend to incubate at either room temperature...

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Q1. Are clear bottomed wells required for plates used in this assay?
A1. Yes

Q2. Serum (1%) will be present in medium when JC10 is added. Will JC10 be effected by serum?
A2. No
Q3. Is there an optimal height setting at which t...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"