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Tetramethylrhodamine methyl ester (perchlorate)

Catalog No.GC19476

Tetramethylrhodamine methyl ester (perchlorate) Chemical Structure

Non-cytotoxic cell-permeant

Size Price Stock Qty
5mg
$48.00
In stock
10mg
$91.00
In stock
25mg
$169.00
In stock
50mg
$266.00
In stock

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Sample solution is provided at 25 µL, 10mM.

Quality Control

Quality Control & SDS

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Protocol

Cell experiment:

Cultures are exposed to Millipore-filtered solutions (0.22 µm) containing TMRM Perchlorate for 1 hr at 37°C (except the experiment involving different durations of exposure to TMRM Perchlorate). After treatment, solutions are removed and growth media reapplied under sterile conditions, and cultures are post-incubated for 18 hours at 37°C (except for the experiment involving analysis at different time points after exposure). Cells are then stained with 2 mg/mL bisbenzimide for 20 min at room temperature. Coverslips are subsequently washed in saline and imaged using 2P microscopy. Apoptotic cells are identified as brightly fluorescent nuclei under UV excitation indicating DNA fragmentation. Cell survivability is calculated as the percentage of live, unstained cells (±SD) in five microscopic fields per treatment[1].

References:

[1]. 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.
[2]. 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.

Background

Tetramethylrhodamine methyl ester (TMRM) (perchlorate) is a non-cytotoxic cell-permeant fluorogenic dye most commonly used to assess mitochondrial function using live cell fluorescence microscopy and flow cytometry.1,2,3 It has two excitation peaks at 515 and 555 nm and an emission peak in the red-orange range (575 nm). Due to the polarization of the mitochondrial membrane, TMRM is taken up into healthy mitochondria. However, when the membrane is depolarized, as in apoptosis, it is not taken up or is released from the mitochondria. Thus, the strength of the fluorescence signal in mitochondria is used to assess cell viability.

Reference: 

1. Farkas, D.L., Wei, M.-d., Febbroriello, P., et al. Simultaneous imaging of cell and mitochondrial membrane potentials Biophys J. 56(6), 1053-1069 (1989).
2. Gandra, P.G., Nogueira, L., and Hogan, M.C. Mitochondrial activation at the onset of contractions in isolated myofibres during successive contractile periods J. Physiol. 590(15), 3597-3609 (2012).
3. Michea, L., Combs, C., Andrews, P., et al. Mitochondrial dysfunction is an early event in high-NaCl-induced apoptosis of mIMCD3 cells Am. J. Physiol. Renal Physiol. 282(6), F981-F990 (2002).

Chemical Properties

Cas No. 115532-50-8 SDF
Synonyms TMRM, TMR methyl ester
Chemical Name 3,6-bis(dimethylamino)-9-[2-(methoxycarbonyl)phenyl]-xanthylium, perchlorate
Canonical SMILES O=C(OC)C(C=CC=C1)=C1C2=C3C=CC(N(C)C)=CC3=[O+]C4=C2C=CC(N(C)C)=C4.O=Cl(=O)([O-])=O
Formula C25H25N2O3 • ClO4 M.Wt 500.9
Solubility DMF: 30 mg/ml,DMSO: 30 mg/ml,Ethanol: slightly soluble,PBS (pH 7.2): slightly soluble Storage Store at -20°C, protect from light
General tips For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.
Shipping Condition Evaluation sample solution : ship with blue ice
All other available size: ship with RT , or blue ice upon request

In vivo Formulation Calculator (Clear solution)

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Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.

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