MitoTools: Mitochondrial research has never been so easy

Involved in many physiological processes (apoptosis, aging, cell signalling..) and numerous diseases (cardiovascular disorders, to cancer and neuro-degenerative diseases…), Mitochondria have become more and more popular as a study model in many research fields. In order to facilitate your daily work, tebu-bio provides a broad range of easy-to-use kits and assays: read on to discover the MitoTools!

Research tools aimed at exploring mitochondria are becoming more and more popular and cover multiple experimental applications. tebu-bio provides a full range of mitochondrial research kits developed by Dojindo, covering many mitochondrial research fields as mentioned below.

Mitochondrial Membrane Potential detection by Fluorescence

Fig.1 : Mitochondrial Membrane detection with JC-1 MitoMP Detection Kit

A decrease in Mitochondrial activity is know to be closely related to cellular dysfunction and many diseases (cancer, Alzheimer’s…). Mitochondria membrane potential can be used as a marker to evaluate this state. Dojindo has developed a fluorescent kit, the JC-1 MitoMP Detection Kit, to measure this potential. In healthy Mitochondria (High Membrane Potential), JC-1 compound will penetrate and form polymer to fluoresce in red (fig. 1). When membrane potential decreases, JC-1 will become a monomer and show green fluorescence.

Fig.2 : JC-1 MitoMP Detection Kit detection with several equipments
Jurkat cells treated by apoptosis inducing reagent, Staurosporine, were stained with JC-1 MitoMP Detection Kit

This ready to use kit contains the JC-1 dye which will be dissolved perfectly in your sample and the imaging buffer. As shown in figure 2, this JC-1 MitoMP detection kit can be used with several types of equipment. Finally, the change in red-green fluorescence ratio is used as an indicator of Mitochondrial condition.

Simplified Mitophagy Detection:

Fig.3 : Mitophagy detection with the Fluorescnet Mitophagy detection kit

Composed of a Mtphagy Dye (detection of Mitophagy), this Mitophagy Detection kit allows simple detection of the Mitophagy events by increase of fluorescence. The Mtphagy dye will accumulate and emit weak fluorescence in intact Mitochondria. After induction of Mitophagy events, the damaged mitochondria fuses with Lysosome, and under acidic condition the Mtphagy dye emits a strong fluorescence (fig.3). A lyso dye is also included in the kit to confirm the fusion of the mitochondria with the Lysosome.

Fig.4 : Mitophagy Detection in starved cells
Living HeLa cells are co-stained with Mtphagy Dye and Lyso Dye under mitophagy induced condition.

Based on a simple 4 steps protocol this Mitophagy detection kit will allow you a strong detection of Mitophagy events (Fig. 4) for all your experimental conditions.

Mitochondrial Iron Detection Reagent

Known as the most abundant transition metal element in the organism, Iron is getting attention because it’s suggested to be related to cellular death and damages. Due to it’s water solubility and the global intracellular reductive environment, understanding the behavior of ferrous ion (Fe 2+) appears as an important point.

Fig.5 : Fe2+ detection in Mitochondria with the Mito-FerroGreen probe
Co-localized Mito-FerroGreen and mitochondrial staining dye, MitoBright Deep Red (Dojindo, Code: MT08) were observed after addition of iron(II) in living HeLa cells.

To specifically detect Fe2+ in living cell, Dojindo have developed a cell permeable and highly specific probe the Mito-FerroGreen for ferrous ion detection in Mitochondria (fig.5).

Mitochondrial Lipophilic Peroxide Detection Reagent

Lipid peroxidation is the metabolic process under the oxidative stress such as the presence of reactive oxygen species (ROS). Lipid peroxide is an important compound which formed by the chain reaction of lipid radicals and is a marker of oxidative damage in organelle membranes.

Fig.6 : Detection of Lipophilic peroxide with the MitoPeDPP

MitoPeDPP is a newly developed, cell permeable fluorescent probe for imaging the lipophilic peroxide in living cell with fluorescent microscopy (fig. 6). After accumulation into the mitochondrial inner membrane, the
MitoPeDPP probe is oxidized by lipophilic peroxide and will emits a strong fluorescence. With excitation and emission wavelengths of the oxidized MitoPeDPP (Ox-MitoPeDPP) of 452 nm and 470 nm, respectively, photodamage and autofluorescence of the samples are minimized.

Mitochondrial Singlet Oxygen Detection Reagent

Fig.7 : Mitochondrial Singlet Oxygen detection by the Si-DMA probe

Composed of silicon-containing rhodamine as fluorophore and an anthracene moieties as active site, the new Si-DMA probe is able to detect specifically (among 7 other ROS) singlet oxygen in Mitochondria (fig.7).

After reaction with Singlet Oxygen, the fluorescence emitted by the Si-DMA will increase about 17 times. In addition, Si-DMA is able to visualize real-time generation of ¹O2 from protoporphyrin IX in mitochondria with 5-aminolevulinic acid (5-ALA), a precursor of heme (fig.8).

Fig.8 : Real time Singlet Oxygen detetion in Mitochondria after 5-ALA addition

Mitochondrial Staining reagents

Dojindo have developed a range of fluorescent probess for selective staining of Mitochondria in living cells: MitoBright.

Fig.9 : MitoBright series for Mitochondrial staining

The three MitoBright probes will accumulate into healthy Mitochondria due to the membrane potential dependency. Thanks to it’s ability to make covalent bond to proteins, MitoBright probes will be well retained in the mitochondria during all your experiments.

Interested in facilitating your Mitochondrial research? Contact your local tebu-bio office to get your quote and to request the full brochure of all our tools to analyse the Cellular Function.

Check all of our other solutions to improve your Mitochondrial research:

Featured image courtesy of Dojindo Laboratories, subject to copyright.

Frédéric Samazan
Written by Frédéric Samazan
Frédéric Samazan is a Product Manager at tebu-bio, and an avid football fan in his spare time.