TMRM

Cat. No.: HY-D0984: FAQs
Data Sheet Handling Instructions

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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.

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TMRM Chemical Structure

CAS No. : 115532-49-5

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Other In-stock Forms of TMRM:

TMRM Perchlorate

Other Forms of TMRM:

TMRM Perchlorate In-stock

14 Publications Citing Use of MCE TMRM

: TMRM purchased from MedChemExpress. Usage Cited in: Endocrinology. 2020 Sep 1;161(9):bqaa119. [Abstract]; Mitochondrial membrane potential detected by TMRE. TMRM staining reveals a decreased signal in the Immp2l knockdown granulosa cells compared to control cells, suggesting mitochondrial membrane potential depolarization. Melatonin treatment prevented the membrane depolarization in Immp2l knockdown granulosa cells.

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Description	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].
In Vitro	1.Preparation of TMRM working solution 1.1Preparation of the stock solution Dissolve 1 mg TMRM in 525 μL DMSO to obtain 5 mM of stock solution. 1.2Preparation of TMRM working solution Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-20 μM of working solution. Note: Please adjust the concentration of TMRM working solution according to the actual situation. 2.Cell staining 2.1 Suspension cells (6-well plate) a.Centrifuge at 1000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×10⁶/mL. b.Add 1 mL of working solution, and then incubate at room temperature for 5-30 minutes. c.Centrifuge at 400 g at 4℃ for 3-4 minutes and then discard the supernatant. d.Wash twice with PBS, 5 minutes each time. e.Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry. 2.2 Adherent cells a.Culture adherent cells on sterile coverslips. b.Remove the coverslip from the medium and aspirate excess medium. c.Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 30-60 minutes. d.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. TMRM Related Antibodies
Molecular Weight	401.48
Formula	C₂₅H₂₅N₂O₃
CAS No.	115532-49-5
Emission (Em)	576
Excitation (Ex)	550
SMILES	O=C(C1=CC=CC=C1C2=C3C=CC(N(C)C)=CC3=[O+]C4=C2C=CC(N(C)C)=C4)OC
Shipping	Room temperature in continental US; may vary elsewhere.
Storage	Please store the product under the recommended conditions in the Certificate of Analysis.
Purity & Documentation	Data Sheet (277 KB) Handling Instructions (2659 KB)
Dyeing Example	Description: TMRM is a rapidly and selectively mitochondrial membrane potential fluorescent probe with red fluorescence. Method: For mitochondrial membrane potential measurement. 1. Cultures are exposed to Millipore-filtered solutions (0.22 µM) containing TMRM and/or drugs for 1 h at 37°C (except the experiment involving different durations of exposure to TMRM). 2. After treatment, solutions are removed and growth media reapplied under sterile conditions, and cultures are post-incubated for 18 hours at 37°C. 3. Use a confocal laser scanning microscope for image. Description: TMRM is a rapidly and selectively mitochondrial membrane potential fluorescent probe with red fluorescence. Method: For mitochondrial membrane potential measurement. 1. Testing cells are pretreated first. 2. Incubate cells with TMRM (20 μM; 30 min; 37°C; dark). 3. Subsequently, the cells are rinsed with PBS for twice and fixed in 10% formaldehyde neutral buffer solution at room temperature for 10 min. 4. Cells are imaged under a Zeiss LSM710 Confocal laser-scanning microscope. Description: TMRM is a rapidly and selectively mitochondrial membrane potential fluorescent probe with red fluorescence. Method: For mitochondrial membrane potential measurement. 1. Testing cells are pretreated first. 2. The TMRM fluorescent dye is formulated into 50 nM/L with PBS. Incubate cells with TMRM (50 μL; 25 min; 37°C; dark). 3. The fluorescence intensity of membrane potential measurement is detected by High Content Cell Imaging Analysis System (IN Cell analyzer 2500HS, GE, USA) and use a confocal laser scanning microscope for image.
References	[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]

Cell Assay
^[1]

Cultures are exposed to Millipore-filtered solutions (0.22 µm) containing TMRM and/or drugs for 1 hr at 37°C (except the experiment involving different durations of exposure to TMRM). 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].

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

References

[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]

TMRM Related Classifications

Inflammation/Immunology

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Help & FAQs

Do most proteins show cross-species activity?
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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TMRM115532-49-5Fluorescent DyeInhibitorinhibitorinhibit

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