JC-1 - Mitochondrial Membrane Potential Assay Kit

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PubMed

Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB113850)

Using JC1 dye to examine mitochondrial membrane potential depolarization in SH-SY5Y cells.

Son MS et al. (2017) used JC1 Mitochondrial Membrane Potential Assay Kit ab113850 to stain :
- untreated SH-SY5Y cells (control),
- SH-SY5Y cells treated with 1mM MPP⁺ (MPP⁺) and,
- SH-SY5Y cells pretreated with BDS-II followed by 1mM MPP⁺ treatment (MPP⁺ +BDSII).

Normal mitochondrial membrane potential is shown in red with JC-1 dimers and depolarized membrane potential is shown in green in JC-1 monomers.

Image courtesy of Son M S et al. Sci Rep. 2017; 7: 2075. doi: 10.1038/s41598-017-02129-w. Reproduced under the Creative Commons License http://creativecommons.org/licenses/by/4.0/.

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PubMed

Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB113850)

JC1 used to assay in vitro metabolic effect on hepatocyte and hepatoma cells.

Koukourakis MI et al. (2016) used the MMP assay kit to stain with JC-1 in NCTC hepatocytes exposed to amifostine (100 μg/ml) over a time course of 20 minutes in vitro. Confocal microscopy used to assess mitochondrial membrane potential in the cells.

Serial images confirmed a rapid drop of both green and red fluorescence, one minute after exposure, an effect that was restored to normal at 20 minutes, after a small period of a rebound increase.

Image courtesy of Koukourakis M I et al. Sci Rep. 2016; 6: 30986doi: 10.1038/srep30986. Reproduced under the Creative Commons License http://creativecommons.org/licenses/by/4.0/

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AbReview

Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB113850)

JC1 Mitochondrial Membrane Potential Assay Kit using green fluorescence imaging.

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.

This image is courtesy of an Abreview submitted by Dimitra Kalamida

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Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB113850)

JC-1 assay result in HL60 cells treated with Troglitazone.

JC1 - Mitochondrial Membrane Potential Assay Kit (ab113850). 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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Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB113850)

JC-1 assay result in HL60 cells treated with FCCP.

JC1 - Mitochondrial Membrane Potential Assay Kit (ab113850). 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.

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Functional Studies - JC-1 - Mitochondrial Membrane Potential Assay Kit (AB113850)

JC-1 assay result in HepG2 cells treated with CCCP.

JC1 - Mitochondrial Membrane Potential Assay Kit (ab113850). 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.