- Catalogue No.
K125
- Size
100T
- Kit component
|
Item
|
Size（100T）
|
Storage
|
|
Assay buffer
|
100 mL*2
|
-20℃
|
|
Calcein AM(1000x)
|
100 μL
|
-20℃, shading light
|
|
Cocl2(100x)
|
1 mL
|
-20℃
|
|
Ionomycin(200x)
|
500 μL
|
-20℃, shading light
|
- Storage
The kit should be stored at -20℃ shading light for 12 months.
- Principle of the Assay
Mitochondrial Permeability Transition Pore Assay Kit or MPTP Assay Kit is a kit that uses the membrane-permeable fluorescent probe Calcein AM to detect the opening degree of the mitochondrial permeability transition pore. It is also often used in the study of cell death such as apoptosis and necrosis. The kit is a more direct detection method for detecting the opening of MPTP than only based on mitochondrial membrane potential analysis.
Firstly, loading Calcein AM through passive transportation, which is a kind of cell staining reagent for fluorescent labeling of living cells. It can easily penetrate the living cell membrane and is cleaved by intracellular esterase to form the membrane-impermeable polar molecule Calcein, which is then trapped in the cell and causes the cytoplasm including mitochondria to emit strong green fluorescence. After adding CoCl2, the fluorescence from the cytoplasm is quenched by CoCl2, only left fluorescence in the mitochondria. As a control, cells can be loaded with Calcein AM and CoCl2, and treated with Ionomycin, so as to make cells load more Ca2+, which causes the activation of MPTP and quenching of mitochondrial fluorescence.
- Additional Materials Required
Cell culture plate, Precision Pipettes, Disposable Pipette Tips, Centrifuge, Fluorescence Microscopy or Flow Cytometer
- Notes
1.This assay kit is for Research Use Only. We will not response for any arising problems or legal responsibilities causing by using the kit for clinical diagnosis or other purpose.
2.Please read the instructions carefully and adjust the instruments before the experiments. Please follow the instructions strictly during the experiments.
3.Protection methods must be taken by wearing a lab coat and latex gloves during the experiment.
4.Fluorescent dyes are all prone to quenching. Please try to avoid light as much as possible to slow down fluorescence quenching.
5.Calcein AM (1000X) should avoid repeated freezing and thawing as much as possible. If used multiple times, please aliquot appropriately.
6.CoCl2 is corrosive, harmful or irritating to the human body. It has specific organ toxicity to the respiratory and reproductive systems, or teratogenic and carcinogenic toxicity. Please handle it with care and ensure effective protection to avoid direct contact with the skin or inhalation. Also, be cautious to prevent corrosion of other items. CoCl2 is toxic or harmful to aquatic organisms and must not be directly discharged into the environment.
