JC1 - Mitochondrial Membrane Potential Assay Kit (ab113850)

製品の概要

  • 製品名JC1 - Mitochondrial Membrane Potential Assay Kit
    Mitochondrial Transmembrane Potential キット 製品一覧
  • 検出方法Fluorescent
  • テスト
    1 x 96 test
  • ステップMultiple steps standard assay
  • 製品の概要

    JC1- Mitochondrial Membrane Potential Assay Kit (ab113850) contains tetraethylbenzimidazolylcarbocyanine iodide (JC-1), a cationic dye that accumulates in energized mitochondria. At low concentrations (due to low mitochondrial membrane potential) JC-1 is predominantly a monomer that yields green fluorescence with emission of 530±15nm. At high concentrations (due to high mitochondrial membrane potential) the dye aggregates yielding a red to orange colored emission (590±17.5nm). Therefore a decrease in the aggregate fluorescent count is indicative of depolarization whereas an increase is indicative of hyperpolarization.

     

     

  • アプリケーション適用あり: Functional Studiesmore details
  • 試験プラットフォームReagents

法規制情報

製品の特性

  • 関連性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.
  • 別名
    • mitochondrial membrane potential

アプリケーション

Our Abpromise guarantee covers the use of ab113850 in the following tested applications.

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

アプリケーション Abreviews 特記事項
Functional Studies Use at an assay dependent concentration.

JC1 - Mitochondrial Membrane Potential Assay Kit 画像

  • HL60 cells were seeded and labeled according to section 11.1 of the protocol. Cells were then treated for 4 hours with 100 µM FCCP or vehicle/diluent control (DMSO).  Mean and standard deviation is plotted for 3 replicates from each condition.

  • HepG2 cells were seeded and labeled according to section 11.2 of the protocol. Cells were then treated for 4 hours with a titration series of CCCP (carbonyl cyanide 3-chlorophenylhydrazone) and both monomer and aggregate forms were read on a Perkin Elmer-Wallac 1420 Victor 2 Multilabel plate reader. Mean and standard deviation of aggregate/monomer ratios is plotted for 12 replicates for each concentration. IC50 of CCCP in HepG2 cells was calculated at 8.7 µM

  • HL60 cells were seeded and labeled according to section 11.1 of the protocol. Cells were then treated for 4 hours with a titration series of the thiazolidinedione Troglitazone and both monomer and aggregate forms were read on a Perkin Elmer-Wallac 1420 Victor 2 Multilabel plate reader. Mean and standard deviation of aggregate/monomer ratios is plotted for 3 replicates for each concentration. IC50 of Troglitazone in HL60 cells was calculated at 1.2 µM

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JC1 - Mitochondrial Membrane Potential Assay Kit (ab113850) 使用論文

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Abreviews
The JC1- Mitochondrial membrane potential assay kit has been tested using HepG2 cells, control cells and FCCP-treated cells (100uM for 4h) have been used as a positive control. The company's instructions were followed for JC1 mitochondrial membrane potential assay. Imaging was performed on a customized Andor Revolution Spinning Disk Confocal System built around a stand (IX81 Olympus) with a 60x lens and a digital camera (Andor Ixon+885) (CIBIT Facility, MBG-DUTH). Image acquisition was performed in Andor IQ 2 software. Optical sections were recorded every 0.3 µm. All confocal microscopy images presented in this work are 2D maximum intensity projections of z-stack images (ImageJ 1.47v National Institute of Health,USA).
Personal feedback: A green laser with the appropriate emission filter (530nm) has been used to detect the monomer of the JC1 dye, following FCCP treatment the mitochondrial membrane potential of the cells was eliminated, as demonstrated by the increase of the monomer emission.
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Dr. Dimitra Kalamida

Verified customer

投稿 Nov 09 2016


The lab confirmed that the protocol is not very clear in that part you pointed out. We will update the protocol shortly.

You should add 100 uL of the 1X buffer to the plate after washing - to prevent the cells from drying out.

Looking at JC1 staining it appears that flow cytometry would be possible to perform as long as the cytometer is capable of the proper excitation/emission required. Protocol booklet mentions on Pg 5 that Excitation = 540 – 590 nm and Emission =...

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If you wanted to keep the relation between JC-1 dye and the detected mitochondrial membrane potential after fixation this is not possible. This is because JC-1 is dependent on mitochondrial membrane potential which will be abolished by the fi...

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Thank you for contacting us. It is fine to go ahead and use media without phenol red instead of buffer. Please let me know if you have any further questions.

The most important thing with the protocol is to read membrane potential in the presence of compound (or treatment). The reason is that we have found in our experience that changes in membrane potential can be reversible as soon as the treatment is rem...

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Thank you for contacting us.


The kit contains enough buffer to perform 100 tests. We give 10mL of buffer at 10X which should give a total of 100mL of buffer at 1X (20mL are used to dilute compounds for toxicity assays, 10mL are used for...

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Thank you for contacting us.

The cost of the JC1 - Mitochondrial Membrane Potential Assay Kit (ab113850) is $245 for 100 tests or in other words, one 96-well plate. For a proforma document stating the quote, we will need your billing informa...

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Thank you for contacting us. We have not tested these kits on yeast samples; however there are papers in the literature showing the use of TMRE in yeast as well as measurement of ATP using luciferin. See below title of paper and methods details. Attent...

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Thank you for your inquiry. I heard back from the lab regardingthese two distinct dyes that both measure mitochondrial membrane potential. TMRE stains mitochondria only when there is a membrane potential. JC-1 is slightly different in that it has dist...

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