Lysosome Function Analysis - Selection Guide for Detection and Imaging Reagent / Probe / Kit

Science Note

[Oct. 1, 2024]                                                                                                                                                                                                                            Previous Science Note
Lysosomal Dysfunction: A Key Factor in Alzheimer's Progression

Some research on Alzheimer's disease (AD) shows that lysosomal dysfunction is observed in the early stages of AD and affects the progression of AD. Here are some of the studies that show one of the mechanisms of lysosomal dysfunction in AD and ideas for therapy.

Lysosomes play a critical role in the removal and degradation of cellular waste, including the degradation of proteins and damaged organelles by autophagy. Lysosomal dysfunction is commonly observed in Alzheimer's disease (AD), contributing to the accumulation of amyloid beta plaques and tau tangles that are hallmarks of the disease. Impaired lysosomal activity disrupts cellular homeostasis, leading to the accumulation of toxic proteins and organelle damage, further exacerbating neurodegeneration. Restoring lysosomal function has emerged as a potential therapeutic target for slowing or ameliorating the progression of Alzheimer's disease.

Accumulation of APP C-terminal fragments causes endolysosomal dysfunction through the dysregulation of late endosome to lysosome-ER contact sites
Click here for the original article:Marine Bretou, et. al.,Developmental Cell, 2024.

Urolithin A improves Alzheimer's disease cognition and restores mitophagy and lysosomal functions
Click here for the original article:Yujun Hou, et. al., Alzheimers Dement., 2024.

Rescue of ApoE4-related lysosomal autophagic failure in Alzheimer’s disease by targeted small molecules
Click here for the original article:Meenakshisundaram Balasubramaniam, et. al., Communications Biology, 2024.

Point of Interest

- Endolysosomal collapse in Alzheimer's disease (AD), initiated by decreased lysosomal calcium and increased cholesterol, is triggered by γ-secretase inhibition, with APP depletion rescuing these dysfunctions.

- APP C-terminal fragments (CTFs) are localized to late endosome-lysosome-ER contacts, and excess APP CTFs cause lysosomal Ca2+ deficits, leading to cholesterol accumulation at endosome-lysosome-ER contacts.

- Failure to maintain balanced APP-CTF levels perpetuates signaling, causing lysosomal dysfunction and contributing to early AD pathology.

Point of Interest

- Long-term treatment with urolithin A (UA) improves learning, memory and olfactory function in Alzheimer's disease mice.

- UA reduces amyloid beta and tau pathology and improves mitophagy by restoring lysosomal function through cathepsin Z regulation.

- UA shows therapeutic potential for Alzheimer's disease by improving lysosomal function and modulating immune responses and Alzheimer's disease-related pathways.

Point of Interest

- The APOE ε4 allele increases the risk of Alzheimer's disease by impairing autophagy by blocking the transcription of autophagy genes.

- Small molecules have been identified that bind ApoE4 and restore autophagy gene transcription, thereby reducing amyloid aggregation in models.

- These findings suggest that ApoE4-targeted drugs could be used to rescue lysosomal autophagy in neurodegenerative diseases such as Alzheimer's.

Related Techniques
         Lysosomal function Lysosomal Acidic pH Detection Kit -Green/Red and Green/Deep Red
         First-time autophagy research Autophagic Flux Assay Kit
         Autophagy detection DAPGreen/ DAPRed (Autophagosome detection), DALGreen (Autolysosome detection)
         Mitophagy  detection Mitophagy Detection Kit
         Mitochondrial membrane potential detection JC-1MitoMPDetection Kit,MT-1MitoMPDetection Kit
         Glycolysis/Oxidative phosphorylation Assay Glycolysis/OXPHOS Assay KitExtracellular OCR Plate Assay Kit
         Oxygen consumption rate assay Extracellular OCR Plate Assay Kit
         Intracellular calcium ion detection Fura 2-AM
         Glutathione Quantification GSSG/GSH Quantification Kit
         Cell Proliferation / Cytotoxicity Assay Cell Counting Kit-8Cytotoxicity LDH Assay Kit-WST
Related Applications

Analysis of autophagic flux without transfection 

DALGreen and DAPRed labeled HeLa cells were used to evaluate changes in autophagic flux induced by the lysosomal acidification inhibitor bafilomycin A1 (Baf. A1). Compared to starvation conditions, the fluorescence signals of DALGreen were decreased under inhibited conditions of autolysosome formation by the addition of Baf. A1. In contrast, the fluorescence signals of DAPRed were increased under the same conditions, indicating that Baf. A1 led to the accumulation of autophagosome.

Experimental Conditions
CTRL: Normal condition, Stv.: Induction of autophagy, Stv. + Baf. A1: Inhibition of autolysosome formation
DALGreen filter set: 488 nm (Ex), 490–550 nm (Em)
DAPRed filter set: 561 nm (Ex), 565–700 nm (Em)

Products in Use
Autophagic Flux Assay Kit

Why is Lysosomal Function Important?

The lysosome has been revealed to be an important organelle with a complex role in nutrient sensing and multifaceted signaling. Its importance has been emphasized in research fields as diverse as a neurological disease, cancer, immunity, and senescence.

Recent research reveals that lysosome acidification declines in neurons well before extracellular amyloid deposition, thus lysosomal function is now a hot topic in Alzheimer's disease research.


Lysosomal pH Detection Dye: pHLys Red
pHLys Red has a high sensitivity to lysosomal pH change with more precise localization, which allows slight pH changes to be detected in physiological conditions like the early phase of the disease.

 

 

Lysosomal Mass Detection Dye: LysoPrime Green, Deep Red
Dojindo’s LysoPrime Green / Deep Red more precisely detects lysosomal mass due to their unique specificity for lysosomes, and retention independent of lysosomal pH. In addition, the retention time of the dyes in lysosomes has been extended, allowing data to be acquired over time.

 

Recommended for First Time Users
Lysosomal Acidic pH Detection Kit
Two types of dyes and lysosomal pH acidification inhibitors are available as a set. Positive control is included for immediate testing in your experimental system.


- Lysosomal mass detecting dye: LysoPrime Green
- Lysosomal pH detection dye: pHLys Red
- Lysosomal pH acidification inhibitor: Bafilomycin A1


Application data

 

Endocytic vesicles were labeled by ECGreen and the lysosomal mass and pH were detected separately with LysoPrime Deep Red and pHLys Red. Co-staining with ECGreen and Lysosomal dyes showed the inhibition of endocytic vesicle-fusion induced by Bafilmycin A1.

 

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 Deep Red - 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.

 

Lysosome Staining Reagents and Kits

Explore Dojindo's wide range of lysosomal staining and pH detection dyes. Choose the following kit or reagent that aligns with your experimental requirements.

Product Name
(Item Code)
Supported Devices Indicator and Detection Color Dyes and
Fluorescence Properties
Approximate Number
of Use
Lysosomal Acidic pH Detection Kit-Green/Deep Red (L268) pH pHLys Green
Ex: 488 nm / Em: 490-550 nm
[for 1 set]
35 mm dish: 10 dishes
μ-Slide 8 well: 10 plates
96-well Plate: 2 plates
quantity LysoPrime Deep Red
Ex: 633 nm / Em: 640-700 nm
Lysosomal Acidic pH Detection Kit-Green/Red (L266) Need G/Y Laser
G:532 nm
Y:561 nm
pH pHLys Red
Ex: 561 nm / Em: 560-650 nm
quantity LysoPrime Green
Ex: 488 nm / Em: 500-600 nm
pHLys Red- Lysosomal Acidic pH Detection  (L265) pH pHLys Red
Ex: 561 nm / Em: 560-650 nm
[for 1 tube]
35 mm dish: 10 dishes
μ-Slide 8 well: 10 plates
96-well Plate: 2 plates
LysoPrime Deep Red - High Specificity and pH Resistance  (L264) quantity LysoPrime Deep Red
Ex: 633 nm / Em: 640-700 nm
LysoPrime Green- High Specificity and pH Resistance  (L261) quantity LysoPrime Green
Ex: 488 nm / Em: 500-600 nm
[for 10 μl]
35 mm dish: 10 dishes
μ-Slide 8 well: 10 plates
96-well Plate: 2 plates

Product Classification

Product Classification