TMRM Perchlorate (solution) 

Rhodamine dyes are membrane-permeable cationic fluorescent probes that specifically recognize mitochondrial membrane potentials, thereby attaching to mitochondria and producing bright fluorescence, and at certain concentrations, rhodamine dyes have low toxicity to cells, so they are commonly used to detect mitochondria in animal cells, plant cells, and microorganisms^[1].
Solvent and concentration: DMSO: 10 mM

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
1. Preparation of TMRM Perchlorate working solution
Dilute the stock solution in serum-free cell culture medium or PBS. The corresponding stock solution can be diluted according to the actual situation. Note that if the solvent is DMSO, the cytotoxicity of DMSO must be considered, and a solvent control should be prepared; if the solvent is pure water, the working solution needs to be filtered and sterilized before adding cells.
Note: Please adjust the concentration of TMRM Perchlorate working solution according to the actual situation.
2.Cell staining
2.1 Suspension cells (6-well plate)
2.1.1 Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×10⁶/mL.
2.1.2 Add 1 mL of working solution, and then incubate at room temperature for 5-30 minutes.
2.1.3 Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
2.1.4 Wash twice with PBS, 5 minutes each time.
2.1.5 Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
2.2.1 Culture adherent cells on sterile coverslips.
2.2.2 Remove the coverslip from the medium and aspirate excess medium.
2.2.3 Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 30-60 minutes.
2.2.4 Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.
Note: If detection by flow cytometry, cells need to be resuspended before staining.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

500.93

C₂₅H₂₅ClN₂O₇

115532-50-8

Liquid

Orange to red

O=C(C1=CC=CC=C1C2=C3C=CC(N(C)C)=CC3=[O+]C4=C2C=CC(N(C)C)=C4)OC.O=Cl(=O)([O-])=O

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Crowley LC, et al. Measuring Mitochondrial Transmembrane Potential by TMRE Staining. Cold Spring Harb Protoc. 2016 Dec 1;2016(12):pdb.prot087361.  [Content Brief]

  [2]. Chowdhury SR, et al. Simultaneous evaluation of substrate-dependent oxygen consumption rates and mitochondrial membrane potential by TMRM and safranin in cortical mitochondria. Biosci Rep. 2015 Dec 8;36(1):e00286.  [Content Brief]

  [3]. Monteith A, et al. Imaging of mitochondrial and non-mitochondrial responses in cultured rat hippocampal neurons exposed to micromolar concentrations of TMRM. PLoS One. 2013;8(3):e58059.  [Content Brief]