The opening of MPTP is a critical event leading to cell death, which was assessed using an MPTP kit (Beyotime Biotech, Nantong, China) following the manufacturer’s protocol [29 (link)]. Briefly, cells on the coverslip were washed and labeled for 30 min in a dark room at 37 degrees and examined under a Leica Thunder DMi8 microscope. Diminished green fluorescence indicates increased MPTP opening.
Mptp kit
Manufactured by Beyotime
Sourced in China
The MPTP kit is a laboratory equipment designed for the analysis and detection of MPTP, a chemical compound. The kit provides the necessary tools and reagents to conduct relevant tests and experiments. Its core function is to facilitate the identification and quantification of MPTP in various samples.
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Lab products found in correlation
4 protocols using mptp kit
1
Assessing MPTP-Induced Mitochondrial Permeability
2
Calcein AM Staining for Cell Viability
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MPTP was determined in accordance with the instructions of the MPTP kit (C2009S, Beyotime). Briefly, the cells were rinsed with PBS, added with an appropriate volume of calcein AM staining solution, fluorescence quenching working solution or Ionomycin control, shaken gently to make the dye evenly cover all cells and incubated at 37°C for 30 min in conditions void of light. Subsequently, the cells were cultured in fresh 37°C preheated complete medium and then incubated at 37°C for 30 min in dark conditions to ensure that the intracellular esterase fully hydrolysed calcein AM to generate calcein with green fluorescence. Next, the cells were rinsed with PBS and observed under a fluorescence microscope (BX63, Olympus) at an excitation wavelength of 494 nm and an emission wavelength of 515 nm.
3
Evaluating Mitochondrial Permeability Transition
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MPTP kit was used for evaluation of mitochondrial permeability transition according to the manufacturer’s instructions (Beyotime, Shanghai, China). A density of 1 × 104 cells in a six–well plate, EtOH–induced LO2 cells were treated with 10 µM of ML334, 10 or 30 µM compound DEA (1 ), DMSO for 8 h, and then pretreated with calcein or calcein and CoCl2. Cells were stained by calcein acetoxymethyl ester (Calcein AM) and MPT function was detected by fluorescence spectroscopy.
4
Mitochondrial Membrane Potential and mPTP Assessment
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MMP was detected with JC-1 (C2003S) and Rhodamine 123 (C2008S) kits purchased from Beyotime Biotechnology (China). JC-1 monomers (green fluorescence) and JC-1 aggregates (red fluorescence) represent a low MMP and high MMP, respectively. Rhodamine 123 shows a green fluorescence, which represents a high MMP, whereas the decrease or disappearance of green fluorescence represents a low MMP.
The opening degree of mPTP was detected by an mPTP kit (C2009S, Beyotime Biotechnology, China). In short, calcein shows a green fluorescence when entering the cell. Once exposed to CO2+, the green fluorescence is quenched. When mPTP is close, calcein is loaded onto the mitochondria and does not make contact with CO2+, resulting in the presentation of green fluorescence. When mPTP is open, calcein flows out of the mitochondria into the cytoplasm and makes contact with CO2+, and the green fluorescence weakens or disappears.
The opening degree of mPTP was detected by an mPTP kit (C2009S, Beyotime Biotechnology, China). In short, calcein shows a green fluorescence when entering the cell. Once exposed to CO2+, the green fluorescence is quenched. When mPTP is close, calcein is loaded onto the mitochondria and does not make contact with CO2+, resulting in the presentation of green fluorescence. When mPTP is open, calcein flows out of the mitochondria into the cytoplasm and makes contact with CO2+, and the green fluorescence weakens or disappears.
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