Mitochondria Research

Science Note

Mitochondrial unfolded protein response in neural stem cell aging 

This article focusing on the mitochondrial protein folding stress is particularly significant in activated neural stem cells (NSCs) and neural progenitor cells (NPCs) within the neurogenic niche. This stress increases with age, leading to dysregulated cell cycles and mitochondrial activity in activated NSCs and NPCs in the dentate gyrus. 

The mitochondrial unfolded protein response regulates hippocampal neural stem cell aging
Chin-Ling Wang, et. al., Cell Metabolism (2023)

Point of Interest
- Elevated mitochondrial protein folding stress in aging neural stem cells
- SIRT7 safeguards neural stem cells by reducing mitochondrial protein folding stress
- SIRT7 overexpression enhances neurogenesis and cognitive function in aged mice
Related Techniques
Glycolysis/Oxidative phosphorylation assay Glycolysis/OXPHOS Assay Kit
Oxygen consumption rate assay Extracellular OCR Plate Assay Kit
Mitochondrial membrane potential detection JC-1 MitoMP Detection Kit / MT-1 MitoMP Detection Kit
Mitochondrial superoxide detection MitoBright ROS - Mitochondrial Superoxide Detection
Mitophagy Detection Mitophagy Detection Kit and Mtphagy Dye
Cellular senescence detection (Live cell imaging or FCM) Cellular Senescence Detection Kit
Cellular senescence detection (Plate reader) Cellular Senescence Plate Assay Kit
Lysosomal function assay Lysosomal pH and mass detection Kit
Related Applications

Metabolic shift to glycolysis in senescenct cells

NAD(+) levels decline during the aging process, causing defects in nuclear and mitochondrial functions and resulting in many age-associated pathologies*. Here, we try to redemonstrate this phenomenon in the doxorubicin (DOX)-induced cellular senescence model with a comprehensive analysis of our products.                       

*S. Imai, et al., Trends Cell Biol, 2014, 24, 464-471

Products in Use
   ① DNA Damage Detection Kit - γH2AX

   ② Cellular Senescence Detection Kit - SPiDER-βGal

   ③ NAD/NADH Assay Kit-WST

   ④ JC-1 MitoMP Detection Kit

   ⑤ Glycolysis/OXPHOS Assay Kit、Lactate Assay Kit-WST

2. Simultaneously detection of Lysosomal and Mitochondrial Dysfunction

We tried the simultaneous detection of lysosomal and mitochondrial dysfunction in Hela cells treated with CCCP or Antimycin (AN). CCCP and AN are well-known inducers of mitochondrial ROS regarding loss of mitochondrial membrane potential. Recent research showed the result that CCCP induces not only mitochondrial ROS but also lysosomal neutralization. To detect mitochondrial ROS, HeLa cells were labeled by MitoBright ROS - Mitochondrial Superoxide Detection, and the lysosomal mass and pH were detected separately with LysoPrime Green and pHLys Red. Co-staining with MitoBright ROS and Lysosomal dyes demonstrated that CCCP causes lysosomal neutralization and mitochondrial ROS induction at the same time.

Products in Use
   - LysoPrime Green
   - pHLys Red
   - MitoBright ROS - Mitochondrial Superoxide Detection

 

Related Product
   - Lysosomal Acidic pH Detection Kit

Mitochondria Research

Mitochondria, which have important functions in cells, are not only a place for ATP production by oxidative phosphorylation, but their activity and dysfunction are closely related to cancer, senescence, and neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. Therefore, understanding the state of mitochondria is particularly important.

 

Mitophagy

Target Mitophagy
Reagent Mitophagy Detection Kit Keima-Red
Priciple 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-sensitive ratiometric fluorescent protein. The excitation spectrum changes according to pH. This protein shows high fluorescence ratio (Ex. 550 nm/440 nm) values in a lysosome.
Accumulating in mitochondria Yes
（Covalent bond）		 -
（Expression protein）
Live-cell staining
（Fixation after live-cell staining）		 ✓
(-)		 ✓
(-)
Fixed cell staining - -
Ex/Em (nm) 530/700  550 & 440/620
Staining time*1 30 min～ *2
Product code MD01 -

*1  Suitable staining time depends on sample type and reagent concentration.
*2  It is necessary to construct a keima expression strain.

Oxidative Stress

  Target Lipophilic Peroxide ROS Singlet oxygen Superoxide
Reagent MitoPeDPP ROS Assay Kit Si-DMA MitoBright ROS
Deep Red		 MitoSOX Red
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  reacts with ROS  generated 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 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 fluorescence.
Accumulating in mitochondria Yes
（Electrostatic interaction）		 - Yes
（Electrostatic interaction）		 Yes
（Electrostatic interaction）		 Yes
（Electrostatic interaction）
Live-cell staining
（Fixed Cell staining）		 ✓
(-)		 ✓
(-)		 ✓
(-)		 ✓
(-)		 ✓
(-)
Fixed cell staining - - - - -
Ex/Em(nm) 452/470 505/525 644/670 540/670 510/580
Common filter set FITC FITC Cy5 TexasRed
Cy5		 TRITC
Staining time*1 30 min～ 15 min～ 45 min～ 10 min~ 10 min～
Product Code M466 R252 MT05 MT16 -

*1  Suitable staining time depends on sample type and reagent concentration.

Mitochondrial Membrane Potential

Target Mitochondrial Membrane Potential
Reagent MT-1 JC-1
Principle Cell-permeant fluorescent dyes 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. A cell-permeant ratiometric fluorescent dye that accumulates in intact mitochondria due to the membrane potential. JC-1 forms aggregate (in healthy mitochondria) with red fluorescence.
As membrane potential decreases, JC-1 becomes monomers, which shows in green fluorescence.
Live-cell staining
（PFA fixation after staing）		 ✓
(✓)		 ✓
(-)
Fixed cell staining - -
Ex/Em(nm) 530-560/570-640 Monomer: 514/529
J-aggregation: 585/590
Common filter set TRITC
TexasRed		 Monomer: FITC
J-aggregation: TRITC; TexasRed
Staining time*1 30  min 10～60  min
Product Code MT13 MT09

*1  Suitable staining time depends on sample type and reagent concentration.

Mitochondria Staining

Target Localization of mitochondria
Reagent MitoBright LT series MitoBright IM Red MitoTracker series Rhodamine 123
Principle

Cell-permeant fluorescent dyes accumulate in intact mitochondria due to the membrane potential.

Cell-permeant fluorescent dyes accumulate in intact mitochondria due to the membrane potential.

Cell-permeant fluorescent dyes accumulate in intact mitochondria due to the membrane potential.

Cell-permeant fluorescent dyes accumulate in intact mitochondria due to the membrane potential.
Live-cell staining
（PFA fixation after staining）		 ✓
(✓)		 ✓
(✓)		 ✓
(✓)		 ✓
(-)
Fixed cell staining - - - -
Combination with immunostaining - ✓ Conditionally -
Long-term retention ✓ × × ×
Ex/Em (nm) Green: 493/508
Red: 547/563
Deep Red:643/663		 549/566 490/516 ～ 644/665 507/529
Common filter set Green: FITC

TRITC

-

FITC
Red: TRITC
DEEP Red: Cy5
Staining time*1 10 min～ 30 min～ 15～45 min 15 min～
Product Code MT10　　MT11　　MT12 MT15 - R233

*1  Suitable staining time depends on the sample type and reagent concentration.

Metal Ion Detection

Target Calcium ion (Ca2+) Iron ion(Fe2+)
Reagent Rhod 2-AM Mito-FerroGreen
Principle A cell-permeant fluorescent probe that accumulates in mitochondria and specifically reacts with calcium ion in mitochondria to emit red fluorescence. A cell-permeant fluorescent probe that accumulates in mitochondria and specifically reacts with ferrous ion in mitochondria to emit green fluorescence.
Accumulating in mitochondria ✓
(Electrostatic interaction)		 ✓
(Electrostatic interaction)
Live-cell staining
（PFA fixation after staing）		 ✓
(-)		 ✓
(-)
Fixed cell staining - -
Ex/Em(nm) 553/576 505/535
Common filter set TRITC FITC
Staining time*1 30～60  min 30 min～
Product Code R002 M489

*1  Suitable staining time depends on the sample type and reagent concentration.

Product Classification

Product Classification

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