2024 Cancer / Neurodegeneration Research Promotion


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Latest Hot Topics and Related Dojindo Products

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Cancer Ferroptosis: Relationship on Metabolism, Lysosome, and Mitochondria

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Ferroptosis is a form of programmed cell death characterized by the accumulation of lipid peroxides to lethal levels and is distinct from other forms of cell death such as apoptosis, necroptosis, and autophagy. In the context of cancer, ferroptosis may act as a tumor suppressor mechanism, as cancer cells often have an increased susceptibility to ferroptosis due to their altered metabolism and increased levels of reactive oxygen species (ROS). Therapeutically, inducing ferroptosis in cancer cells has emerged as a promising strategy for cancer treatment, particularly for tumors that are resistant to traditional therapies such as chemotherapy and radiation. In addition, understanding the specific vulnerabilities of cancer cells to ferroptosis may aid in the design of targeted therapies that exploit these weaknesses, providing a potential avenue to overcome drug resistance and improve patient outcomes.

Dietary restriction of cysteine and methionine sensitizes gliomas to ferroptosis and induces alterations in energetic metabolism
Click here for the original article: Pavan S. Upadhyayula, et. al., Nature Communications, 2023.

Lysosomal cystine governs ferroptosis sensitivity in cancer via cysteine stress response
Click here for the original article: Robert V. Swanda et. al., Molecular Cell, 2023.

Mitochondria regulate intracellular coenzyme Q transport and ferroptotic resistance via STARD7
Click here for the original article: Soni Deshwal et. al., Nature Cell Biology, 2023.

Point of Interest
- Cysteine and methionine deprivation (CMD) can synergize with the GPX4 inhibitor RSL3 to increase ferroptotic cell death and lipid peroxidation.

- A cysteine-depleted, methionine-restricted diet can improve the therapeutic response to RSL3 and prolong survival in a syngeneic orthotopic murine glioma model.

- This CMD diet profoundly alters in vivo metabolome, proteome, and lipidome.

Point of Interest
- Depletion of cysteine induces adaptive ATF4 expression at the transcriptional level.

- A shortage of cystine in lysosomes stimulates ATF4 expression through the AhR signaling pathway.

- A weakened cysteine stress response increases sensitivity to ferroptosis during cysteine deprivation.

- CysRx promotes cancer cell ferroptosis through intracellular nutrient reprogramming.

Point of Interest
- The rhomboid protease PARL cleaves STARD7, allowing its dual localization to the mitochondrial intermembrane space and cytosol.

- The mitochondrial STARD7 supports coenzyme Q synthesis, promotes oxidative phosphorylation, and maintains cristae morphology.

- Its cytosolic counterpart facilitates the transport of coenzyme Q to the plasma membrane and protects against ferroptosis.

- Overexpression of cytosolic STARD7 increases the resistance of cells to ferroptosis and reduces the availability of coenzyme Q in the mitochondria.

Related Techniques
Intracellular / mitochondrial lipid peroxidation detection LiperfluoMitoPeDPP
Intracellular / mitochondrial ferrous ion (Fe2+) detection FerroOrangeMito-FerroGreen
Mitochondrial superoxide detection MitoBright ROS Deep Red - Mitochondrial Superoxide Detection
Oxygen consumption rate assay Extracellular OCR Plate Assay Kit
Lysosomal function Lysosomal Acidic pH Detection Kit-Green/Red and Green/Deep Red NEW
Glycolysis/Oxidative phosphorylation Assay Glycolysis/OXPHOS Assay Kit 
Related Applications

The simultaneous detection of lysosomal function with Mitochondrial ROS and intracellular Fe2+

Lysosomal Function and Iron Homeostasis



Recent reports suggest that lysosomal neutralization can result in iron depletion, consequently leading to the disruption of cell viability. To verify this, HeLa cells were labeled with FerroOrange for Fe2+ detection, and the lysosomal mass and pH were separately detected with LysoPrime DeepRed and pHLys Green (a product currently under development). Co-staining with FerroOrange and Lysosomal dyes demonstrated that Bafilomycin A1 (Baf. A1), an inhibitor of lysosomal acidification, causes iron depletion consistent with the findings reported in the article. Interestingly, the iron chelator, Deferiprone (DFP), did not impact lysosomal pH, suggesting that lysosomal function plays a key role in managing iron homeostasis.

Reference: Ross A Weber, et. al., Mol Cell (2020)

Products in Use
   - FerroOrange
   - pHLys Green*
   - LysoPrime Deep Red

*pHLys Green is available as the "Lysosomal Acidic pH Detection Kit-Green/Deep Red". 

Induction of Ferroptosis by Erastin

Erastin is a known inducer of ferroptosis. By inhibiting the cystine transporter (xCT), erastin inhibits the uptake of cystine. Cystine is the raw material for GSH. Therefore, Erastin ultimately decreases the amount of GSH. Decreased GSH then results in lipid peroxide accumulation and induction of ferroptosis.
The following experimental examples show changes in each aforementioned index as a consequence of erastin stimulation. Measurements are made using Dojindo reagents.

Using erastin-treated A549 cells, we measured intracellular Fe2+, ROS, lipid peroxide, glutathione, glutamate release into the extracellular space, and cystine uptake. As a result, inhibition of xCT by elastin was observed and also the release of glutamate and uptake of cystine were decreased. Furthermore, elastin treatment decreased intracellular glutathione while it increased intracellular Fe2+ , ROS, and lipid peroxides.

①Cystine Uptake        

Cystine Uptake Assay Kit

②Released Glu            

Glutamate Assay Kit-WST

③Intracellular GSH     

GSSG/GSH Quantification Kit

④Intracellular Fe2+     


⑤Intracellular ROS     

Highly Sensitive DCFH-DA

⑥Intracellular Lipid   







Microglial and Neuronal Autophagy: Regulation and Relation to Neurodegenerative Diseases

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Autophagy is a cellular process that degrades and recycles damaged organelles, misfolded proteins, and other cellular debris, and is critical for maintaining cellular homeostasis. Dysregulated autophagy is often observed in the context of neurodegeneration, where either excessive or insufficient autophagic activity can contribute to the accumulation of toxic proteins and organelle dysfunction that are hallmarks of neurodegenerative diseases. Enhancing autophagy has been shown to alleviate symptoms and pathology in models of diseases such as Alzheimer's, Parkinson's, and Huntington's, suggesting a protective role against neurodegeneration. Conversely, excessive autophagy can lead to cell death, suggesting that a delicate balance is required for neuronal health and highlighting the complexity of targeting autophagy as a therapeutic strategy for neurodegenerative diseases.

Autophagy enables microglia to engage amyloid plaques and prevents microglial senescence
Click here for the original article: Insup Choi, et. al., Nature Cell Biology, 2023.
Microglial-to-neuronal CCR5 signaling regulates autophagy in neurodegeneration
Click here for the original article: Beatrice Paola Festa, et. al., Neuron, 2023.
Mitochondrial control of microglial phagocytosis by the translocator protein and hexokinase 2 in Alzheimer’s disease
Click here for the original article: Lauren H. Fairley, et. al., PNAS, 2022.

Point of Interest
- Inhibiting microglial autophagy leads to the disengagement of microglia from amyloid plaques, exacerbating neuropathology in mice with Alzheimer's Disease (AD).

- An autophagy deficiency fosters the development of senescent microglia.

- Pharmacological removal of autophagy-deficient senescent microglia can alleviate neuropathology in mice with AD.

Point of Interest
- Activated microglia mediate non-cell-autonomous inhibition of neuronal autophagy.

- CCL-5, CCL-4, and CCL-3 derived from microglia activate the neuronal CCR5 receptor, leading to the inhibition of neuronal autophagy.

- Levels of CCR5, CCL-3, CCL-4, and CCL-5 are elevated in the brains of mice with Huntington’s disease and tauopathy.

- Inhibiting CCR5, either pharmacologically or genetically, ameliorates neurodegeneration in mice.

Point of Interest
- The translocator protein (TSPO) is pivotal for respiratory metabolism and energy supply in microglia.

- Hexokinase-2 (HK) affects glycolytic metabolism and phagocytosis through its interaction with mitochondria.

- Mitochondrial HK influences glycolysis and inflammation, and its displacement improves phagocytosis in TSPO-deficient microglia.

- Alzheimer’s beta-amyloid drastically stimulated mitochondrial HK recruitment in cultured microglia, which may contribute to microglial dysfunction in Alzheimer’s disease.

Reported Examples: Autophagy and Mitophagy Detection Probes
Detection (Reagent) Samples Reference
 (Autophagic Flux Assay Kit)
Immortalized Fischer rat Schwann cells International Journal of Molecular Sciences


Cortical neurons Neurobiology of Disease
PC12 cells(a mixture of neuroblastic cells and eosinophilic cells) Hindawi
(DAPGreen or DAPRed)
Neurons from mouse embryonic stem cells Life Science Alliance
Hippocampal neurons PNAS
N2a cells (Mouse Albino neuroblastoma) International Journal of Molecular Sciences
(Mtphagy Dye)
Neural progenitor cell (NPC) International Journal of Molecular Sciences
Primary cortical neuronal (CxN) cells Cells
Brain slices (mice) The FEBS Journal

<Cellular Landscape: autophagy-related pathway>


Link to each product (red at left figure)

Autophagy Detection
 Autophagic Flux Assay Kit - First-choice product
 DAPRed - Autophagosome detection
 DAPGreen - Autophagosome detection
 DALGreen - Autolysosome detection

Mitophagy Detection
 Mitophagy Detection Kit and Mtphagy Dye

Mitochondrial Membrane Potential Detection
 JC-1 / MT-1

Lysosomal Analysis
 Lysosomal Acidic pH Detection Kit
                   Green/RedGreen/Deep Red

Endocytosis Detection
 ECGreen-Endocytosis Detection
 AcidSensor Labeling Kit

 Exosome Isolation Kit
 Exosome Membrane Labeling Kit
                          Green / Red / Deep Red






Mitochondrial and Lysosomal, and Iron Regulation of Senescence

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Senescence is a cellular process that results in the cessation of cell division, often serving as a protective mechanism against the proliferation of damaged cells, including potential cancer cells. This process is intricately regulated by numerous factors including, but not limited to, tumor suppressor genes, DNA damage response (DDR) pathways, and various signaling molecules. In addition, the senescence-associated secretory phenotype (SASP), consisting of cytokines, growth factors, and proteases, is regulated by NF-κB and other transcription factors that influence the tissue microenvironment and impact aging and disease processes.

HKDC1, a target of TFEB, is essential to maintain both mitochondrial and lysosomal homeostasis, preventing cellular senescence
Click here for the original article: Mengying Cui, et. al., PNAS, 2023.

Iron accumulation drives fibrosis, senescence and the senescence-associated secretory phenotype
Click here for the original article: Mate Maus, et. al., Nature, 2023.
Microautophagy regulated by STK38 and GABARAPs is essential to repair lysosomes and prevent aging
Click here for the original article: Monami Ogura, et. al., EMBP Reports, 2023.

Point of Interest
- HKDC1, a protein involved in glycolysis, is a direct target of the transcription factor TFEB and is essential for maintaining both mitochondrial and lysosomal function.

- This activity helps avert cellular senescence, playing a vital role in maintaining cellular homeostasis.

- Beyond its role in glycolysis, HKDC1 contributes to mitophagy and lysosomal repair processes independently.

- The absence of HKDC1 may result in cellular senescence and the buildup of damaged organelles.


Point of Interest
- Vascular and hemolytic injury trigger iron accumulation, which causes senescence and promotes fibrosis.

- Senescent cells persistently accumulate iron, even after the increase in extracellular iron has subsided.

- Cells exposed to various types of senescence-inducing insults accumulate abundant ferritin-bound iron, mostly within lysosomes.
- The high levels of labile iron fuel the generation of reactive oxygen species and the SASP.  

Point of Interest
- Microautophagy in the repair of damaged lysosomes prevents aging.

- STK38 and GABARAPs are key regulators of this process.

- STK38 is required for lysosomal recruitment of VPS4 and GABARAPs are involved in ESCRT assembly.

- Depletion of these regulators leads to accelerated cellular senescence and shortened lifespan.

Related Techniques
           Cellular senescence detection SPiDER-βGal for live-cell imaging or flow cytometry / microplate reader / tissue samples.
           First-time autophagy research Autophagic Flux Assay Kit
           Autophagy detection DAPGreen / DAPRed (Autophagosome detection), DALGreen (Autolysosome detection) 
           Lysosomal function Lysosomal Acidic pH Detection Kit-Green/Red and Green/Deep Red NEW
           Ferrous ion (Fe2+) detection FerroOrange(intracellular), Mito-FerroGreen(mitochondria)
           Mitochondrial superoxide detection MitoBright ROS Deep Red - Mitochondrial Superoxide Detection
           Oxygen consumption rate assay Extracellular OCR Plate Assay Kit
Related Applications

Analysis of Lysosomal Mass and pH Exchange in Senescence-induced Cells

Purpose: To investigate changes in lysosomal mass and pH in A549 cells induced to senescence by treatment with Doxorubicin (DOX).

Methods: Senescence-associated acidic β-galactosidase (SA-βGal) activity was detected using Cellular Senescence Detection Kit - SPiDER-βGal. Lysosomal mass was detected using LysoPrime Deep Red, and pH was detected using pHLys Red. Fluorescence imaging was used to observe changes in lysosomal mass and pH in senescent cells compared to non-senescent cells. The normalized fluorescence intensity of lysosomal mass and pH was also measured by a plate reader.

Results: Our findings indicate that senescence induced by DOX resulted in an increase in lysosomal mass and acidification of pH compared to non-senescent cells. The obtained results are consistent with previous reports* that demonstrated enhanced lysosomal activity in senescent cells induced by the CDK4/6 inhibitor, palbociclib. The fluorescence imaging and plate reader data both support these findings.

Miguel Rovira, et. al., Aging Cell (2022)

<Experimental Conditions for Microscopy>
SA-βGal(Green):Ex = 488 nm, Em = 490 – 550 nm
Lysosomal pH (Red):Ex = 561 nm, Em = 560 – 620 nm
Lysosomal mass (Deep Red):Ex = 633 nm, Em = 640 – 700 nm

<Experimental Conditions for Plate Reader>
SA-βGal: Ex = 525 – 535 nm, Em = 550 – 570 nm
Lysosomal pH: Ex = 555 – 565 nm, Em = 590 – 610 nm
Lysosomal mass: Ex = 645 – 655 nm, Em = 690 – 710 nm

<Products in Use>
Cellular Senescence Detection Kit
Lysosomal pH and mass detection Kit
   > More about Lysosomal Function Analysis









Hot Indicators

Trends in various indices of cell function analysis in terms of number of papers (referenced from Dimensions)

Ranked Reagents

Dojindo categories and products are ranked based on sales, online searches, and inquiries over the past six months.


Ranking Category Product (Catalog Page#)


FerroOrange (14), Liperfluo (12), Cystine Uptake Assay Kit (21)



DAPRed, DALGreen, Autophagy Flux Assay Kit (8)



Glycolysis/OXPHOS Assay Kit (19)

4 Senescence Cellular Senescence Detection Kit – SPiDER-ßGal (6)


Mitophagy Detection Kit (26), MitoBright ROS Deep Red (27)



Lysosomal Acidic pH Detection Kit Green/Red (31)



ROS Assay Kit - Highly Sensitive DCFH-DA – (10)



Lipid Droplet Assay Kit – Blue / Deep Red (35)

Promotional Products

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Category Product Name Feature
Cell Proliferation / Cytotoxicity Assay Cell Counting Kit-8 Best Seller Simple-process cell viability assay​ using metabolism (dehydrogenase activity) as an indicator
Cytotoxicity LDH Assay Kit-WST Reliable cytotoxicity assay by released LDH activity
Cell Count Normalization Kit Normalization of cell counts by nuclear staining
Viability/Cytotoxicity Multiplex Assay Kit NEW Set of Cell Counting Kit-8 and Cytotoxicity LDH Assay Kit-WST (500 tests each)
Senescence Cellular Senescence Detection Kit – SPiDER-ßGal HOT First choice for cellular senescence assay
SPiDER-ßGal SA-β-gal assay for living cells and tissue
Cellular Senescence Plate Assay Kit – SPiDER-ßGal Cellular senescence assay with a plate reader
Nucleolus Bright Green Nucleolus fluorescent staining
Nucleolus Bright Red
Ferroptosis Liperfluo HOT Intracellular lipid peroxide measurement
MitoPeDPP Mitochondria lipid peroxidation measurement
Mito-FerroGreen Mitochondria ferrous ion (Fe2+) detection
FerroOrange HOT Intracellular ferrous ion (Fe2+) detection
Lipid Peroxidation Probe -BDP 581/591 C11- Lipid peroxidation detection
ROS / Oxidative Stress GSSG/GSH Quantification Kit Quantification of reduced and oxidized glutathione
ROS Assay Kit - Highly Sensitive DCFH-DA - NEW Higher sensitivity than conventional DCFH-DA
ROS Assay Kit - Photo-oxidation Resistant DCFH- DA - NEW Reduced photo-oxidation and compatible with fixation
MitoBright ROS Deep Red - Mitochondrial Superoxide Detection NEW Mitochondrial superoxide detection by deep red staining, co-staining with other markers
Lysosome Lysosomal Acidic pH Detection Kit-Green/Red NEW Ready-to-use kits for Lysosomal pH and mass detection
Lysosomal Acidic pH Detection Kit-Green/Deep Red NEW
pHLys Red - Lysosomal Acidic pH Detection NEW Highly sensitive lysosomal pH detection dye
LysoPrime Green / Deep Red – High Specificity and pH Resistance NEW Accurate lysosome detection with high selectivity and resistance to pH changes
Cellular Metabolism ATP Assay Kit-Luminescence Easy quantification of intracellular ATP with a plate reader
Glucose Assay Kit-WST (Colorimetric) Easy quantification of various intracellular or extracellular indicators with a plate reader
Glutamine Assay Kit-WST (Colorimetric)
Glutamate Assay Kit-WST (Colorimetric)
Glycolysis/OXPHOS Assay Kit NEW
Glycolysis/JC-1 MitoMP Assay Kit NEW
Lactate Assay Kit-WST (Colorimetric)
α-Ketoglutarate Assay Kit-Fluorometric Easy quantification of various intracellular indicators with a plate reader
NAD/NADH Assay Kit-WST (Colorimetric)
NADP/NADPH Assay Kit-WST (Colorimetric)
Extracellular OCR Plate Assay Kit HOT NEW Oxygen Consumption Rate (OCR) detection with a fluorescent plate reader
Nutrient Uptake Glucose Uptake Assay Kit- Blue / Green / Red Highly sensitive and easy to measure each indicator with a fluorescent plate reader without RI-labeled
Amino Acid Uptake Assay Kit NEW
Cystine Uptake Assay Kit NEW
Fatty Acid Uptake Assay Kit NEW
Mitochondrial Staining MitoBright LT Green / Red / Deep Red Mitochondria imaging with long-term visualization
MitoBright IM Red for Immunostaining Mitochondria imaging with capable of co-staining with immunomarkers
mtSOX Deep Red – Mitochondrial Superoxide Detection NEW Mitochondrial superoxide detection by deep red staining, co-staining with other markers
MitoPeDPP Mitochondria lipid peroxidation measurement
Mito-FerroGreen Mitochondria ferrous ion (Fe2+) detection
Mitochondrial Membrane Potential JC-1 MitoMP Detection Kit Widely used and many papers reported
MT-1 MitoMP Detection Kit NEW More sensitive than JC-1 and fixable after staining
Mitophagy Mitophagy Detection Kit First choice for mitophagy detection
Mtphagy Dye For experienced users of the mitophagy detection kit
Lipid Droplet Lipi-Blue / Green / Red / Deep Red Accurate lipid droplet imaging using living and PFA fixed cells with low background
Lipid Droplet Assay Kit-Blue / Deep Red Accurate lipid droplet measurement with plate reader and flow cytometer
Autophagy DALGreen - Autophagy Detection HOT Selective autolysosome detection in live cells
DAPGreen - Autophagy Detection HOT Selective autophagosome detection in live cells
DAPRed - Autophagy Detection HOT
Autophagy Flux Assay Kit  NEW First-choice product for autophagy research
Endocytosis ECGreen-Endocytosis Detection NEW Accurate detection of endocytosis by pH changes
AcidSensor Labeling Kit – Endocytic Internalization Assay NEW Labeling pH-sensitive fluorescent dye to antibody or protein to monitor the endocytosis uptake
Phagocytosis AcidSensor Labeling Kit – Endocytic Internalization Assay Detect phagocytosis activity with flow cytometry. The application is available on the product page.
Exosome ExoIsolator Exosome Isolation Kit NEW Exosomes are obtained quickly without any complicated operations from a supernatant
ExoIsolator Isolation Filter NEW
ExoSparkler Exosome Membrane Labeling Kit-Green / Red / Deep Red Exosome membrane – fluorescent labeling (highly precise)
Plasma Membrane PlasMem Bright Green / Red Cell membrane staining reagents with low cytotoxicity



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