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

Overview

  • Product nameJC-10 Mitochondrial Membrane Potential Assay Kit (Microplate)See all Mitochondrial Transmembrane Potential kits ...
  • Tests
    5 x 96 well plate
  • Sample type
    Adherent cells, Suspension cells
  • Assay typeDirect
  • Product overview

    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.

    ab112134 Mitochondria Membrane Potential Assay Kit 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.

    ab112134 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.

    Visit our FAQs page for tips and troubleshooting.

  • Tested applicationsFunctional Studiesmore details

Properties

  • RelevanceMitochondrial 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.
  • Alternative names
    • mitochondrial membrane potential

Applications

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.

Application Abreviews Notes
Functional Studies Use at an assay dependent concentration.

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

  • Camptothecin-induced mitochondria membrane potential changes with JC-10 and JC-1 in Jurkat cells. After Jurkat cells were treated with camptothecin (10 µM) for 4 hours, JC-1 and JC-10 dye loading solutions were added to the wells 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.

Protocols

References for JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate) (ab112134)

This product has been referenced in:
  • He Q  et al. Rotenone induces reductive stress and triacylglycerol deposition in C2C12 cells. Int J Biochem Cell Biol 45:2749-55 (2013). Functional Studies . Read more (PubMed: 24104397) »

See 1 Publication for this product

Product Wall

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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Thank you for your call the other day.

The laboratory has not tested the possiblity of fixing the cells after the assay. The assay has not been made for this, so we are really not sure whether this could work. I am very sorry for not being ab...

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Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guar...

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