Mitochondria Research

Why is Mitochondrial Research Important?

Mitochondrial analysis is essential for understanding cellular energy metabolism because mitochondria regulate ATP production, redox balance, and signaling pathways. Thus, assessment of mitochondrial functions such as membrane potential, mitophagy, and respiration provides insight into cellular abnormalities, and these studies are critical in neurodegenerative diseases, cancer biology, and metabolic disorders, helping to uncover therapeutic targets and disease mechanisms.

Selection Guide for Mitochondria-related Reagents

Mitochondria research is very multi-faceted, because the multi-functional organelle is not only involved in energy production in a cell, but other additional cellular functions. The active cycle of mitochondrial fusion and division induces morphological changes, which is called mitochondrial dynamics. Abnormalities in morphological control of mitochondria are associated with neurodegenerative diseases, metabolic disorders, aging, and so on. Therefore, the demand for long-term observation of mitochondrial dynamics has recently been increasing.

Selection Guide of Reagents

The following table lists reagents for mitochondrial research designed to stain and detect mitochondria (MitoBright LT, MitoTracker, etc.), mitochondrial membrane potential (JC-1, TMRM, TMRE, etc.), reactive oxygen species AKA ‘ROS’ (MitoBright ROS, MitoSOX, etc.), mitophagy, and lipid peroxides.

Mitophagy

Mitophagy
Reagent Mtphagy Dye Keima-Red
Principle Mtphagy Dye (included in Mitophagy Detection Kit) is a pH-sensitive fluorescent probe that accumulates in mitochondria and emits red fluorescence due to acidic conditions in a lysosome. A pH-sentitive ratiometric fluorescent protein. The excitation spectrum changes accoring to pH. This protein shows high fluorescence ratio (Ex. 550 nm/440 nm) values in a lysozome.
Fixed cell staining
Live-cell staining Yes Yes
Fixation after live-cell staining
Staining time > 30 min
Ex / Em 530 / 700 440, 550 / 620
Product code MD01MT02

Lipophilic peroxide / Singlet oxygen / Superoxide

  Lipophilic peroxide Singlet oxygen Superoxide Superoxide
Reagent MitoPeDPP Si-DMA MitoBright ROS Deep Red MitoSOX
Principle A cell-permeant fluorescent probe that accumulates in mitochondria and specifically reacts with lipophilic peroxides in mitochondria to emit fluorescence. A cell-permeant fluorescent probe that accumulates in mitochondria and specifically reacts with singlet oxigen generated in mitochondria to emit red fluorescence. A cell-permeant fluorescent probe that accumulates in mitochondria and reacts with superoxide generated in mitochondria to emit fluorescence. A cell-permeant fluorescent probe that accumulates in mitochondria and reacts with superoxide generated in mitochondria to emit red fluorescence.
Fixed cell staining
Live-cell staining Yes Yes Yes Yes
Fixation after live-cell staining
Staining time > 15 min > 45 min > 10 min > 10 min
Ex / Em 452 / 470 644 / 670 540 / 670 510 / 590
Product code M466 MT05 MT16

Membrane potential

Membrane potential
Reagent JC-1 MT-1 TMRM, TMRE
Principle A cell-permeant ratiometric fluorescent dye that accumulates in intact mitochondria due to the membrane potential. The excitation spectrum changes according to the mitochondria membrane potential. Cell-permeant fluorescent dyes that accumulate in intact mitochondria due to the membrane potential. MT-1 is extremely photostable and more sensitive than JC-1 and can provide equivalent detection sensitivity to TMRE. Cell-permeant fluorescent dyes that accumulate in intact mitochondria due to the membrane potential. Diffusion of the probes occurs in a damaged mitochondria that has the decreased membrane potential.
Fixed cell staining
Live-cell staining Yes Yes Yes
Fixation after live-cell staining Yes
Staining time 30-60 min 30 min 30-60 min
Ex / Em Monomer: 514 / 529
J-aggregation: 585/590
530-560 / 570-640 550 / 575
Product code MT09 MT13

Mitochondria staining

Mitochondria staining
Reagent MitoBright LT series MitoBright IM Red MitoTracker series Rhodamine 123
Principle Cell-permeant fluorescent dyes that accumulate in intact mitochondria due to the membrane potential. Cell-permeant fluorescent dyes that accumulate in intact mitochondria due to the membrane potential and covalently binds to proteins and other biomolecules. Cell-permeant fluorescent dyes that accumulate in intact mitochondria due to the membrane potential. Cell-permeant fluorescent dye that accumulates in intact mitochondria due to the membrane potential.
Fixed cell staining
Live-cell staining Yes Yes Yes Yes
Fixation after live-cell staining Yes Yes
Staining time 30 min 30 min 15-45 min > 15 min
Ex / Em 493 / 508, 547 / 563, 643 / 663 548 / 566 490 / 516 ~
644 / 665
507 / 529
Product code MT10, MT11,MT12 MT15 R233

Metal Ion Detection

   Iron ion (Fe2+) Calcium ion (Ca2+)
Reagent Mito-FerroGreen Rhod2-AM
Principle A cell-permeant fluorescent probe that accumulates in mitochondria and specifically reacts with ferrous ion in mitochondria to emit green fluorescence. A cell-permeant fluorescent probe that accumulates in mitochondria and specifically reacts with calcium ion in mitochondria to emit red fluorescence.
Fixed cell staining
Live-cell staining Yes Yes
Fixation after live-cell staining
Staining time 30 min 30-60 min
Ex / Em 505 / 535 553 / 576
Product code M489 R002 
Application Products
Mitophagy Detection Mitophagy Detection Kit
Mitochondrial Phospholipid Peroxidase Detection MitoPeDPP
Mitochondrial Ferrous Ion Detection Mito-FerroGreen
Mitochondrial Superoxide MitoBright ROS - Mitochondrial Superoxide Detection
Mitochondrial Singlet Oxygen Detection Si-DMA for Mitochondrial Singlet Oxygen Imaging
Mitochondrial Membrane Potential JC-1 MitoMP Detection Kit
MT-1 MitoMP Detection Kit
Mitochondria Staining MitoBright LT Green
MitoBright LT Red
MitoBright LT Deep Red
MitoBright IM Red for Immunostaining

Cancer, Cell Death, and Mitochondria

Apoptosis is a protective defense mechanism that effectively suppresses tumor growth and eliminates tumor cells.
One of the main mechanisms that trigger apoptosis is the increase in mitochondrial metabolic activity, which leads to elevated ROS levels in cancer cells. Excessive ROS damage mitochondrial function, causing mitochondrial membrane depolarization, which subsequently activates the intrinsic apoptosis pathway. Tumor cell immune evasion is a key feature of tumor pathophysiology, and mitochondria play a central role in both inhibiting and promoting immune evasion within the  complex mechanism    s involved in immune responses.1)

The Potential of Ferroptosis in Cancer Therapy: Many studies have found that ferroptosis sensitivity can be used to target tumors resistant to conventional therapies (such as triple-negative breast cancer and glioblastoma).2)

Ferroptosis and the Immune Microenvironment: Neutrophils in the tumour microenvironment die spontaneously by ferotosis and the lipid peroxide released suppresses T-cell activity, thereby suppressing tumour immunity.3)

Reference
1) Gao, J., Cancer Gene Therapy, 2024, 31, 970-983
2) Yang, F., Cell Metabolism, 2023, 35(1), 84-100
3) Kim, R., Nature, 2022, 612, 338-346


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Mitochondrial Function and Cellular Senescence

In senescent cells, due to mitochondrial dysfunction, ATP is primarily generated through the anaerobic glycolysis pathway, leading to an increase in lactate production2). DNA damage is one of the causes of mitochondrial dysfunction in cellular aging. The accumulation of DNA damage activates DNA repair mechanisms and increases NAD+ consumption. The decrease in NAD+ levels reduces SIRT1 activity, an important factor in maintaining mitochondrial function, leading to impaired mitochondrial function (inhibition of electron transfer → ATP production / reduction of NAD+ levels)1),3).

Reference:

1. J. Wu, Z. Jin, H. Zheng and L. Yan, “Sources and implications of NADH/NAD+redox imbalance in diabetes and its complications”, Diabetes Metab. Syndr. Obes., 2016, 9, 145

2. Z. Feng, R. W. Hanson, N. A. Berger and A. Trubitsyn, “Reprogramming of energy metabolism as a driver of aging”, Oncotarget., 2016, 7(13), 15410.

3. S. Imai and L. Guarente, “NAD+ and sirtuins in aging and disease”, Trends in Cell Biology, 2014, 24(8), 464.

Neurodegeneration-Autophagy and Mitochondrial Damage

Indicator Mitophagy Mitochondrial fission abnormalities ROS MPTP (Mitochondrial Permeability Transition Pore) MMP (Mitochondrial Membrane Potential) ATP
Detection Kit MD01 Mitophagy Detection Kit MT10/MT11/MT12 MitoBright LT Green/Red/Deep Red R252/R253
ROS Assay Kit
- MT09:JC-1 MT13:MT-1 A550 ATP Assay Kit-Luminescence
Alzheimer's disease (AD)
Parkinson's disease (PD) - -
Huntington's disease (HD) - - -
Amyotrophic lateral sclerosis (ALS) -

Refernce :

Role of Mitochondria in NeurodegenerativeDiseases: From an Epigenetic Perspective
 

Mitophagy / Autophagy Analysis Products

Product Name Probe 1
Dyes and Fluorescence Properties
Probe 2
Dyes and Fluorescence Properties
Mitophagy Detection Kit Mtphagy Dye
Ex: 500-560 nm, Em: 670-730 nm
Lyso Dye
Ex: 350-450 nm, Em: 500-560 nm
Mtphagy Dye Mtphagy Dye
Ex: 500-560 nm / Em: 690-750 nm
 
Autophagic Flux Assay Kit* DAPRed
Ex: 500-560 nm / Em: 690-750 nm
DALGreen
Ex: 350-450 nm / Em: 500-560 nm

   *Autophagic Flux Assay Kit includes lysosome acidification Inhibitor as a negative control.

 

Product Classification

Product Classification