Home Transmembrane Transporters ATPase inhibitor - Dyes chemical Rhodamine 123 For research use only.

Rhodamine 123

Synonyms: RH-123, R-22420

Rhodamine 123 (RH-123, R-22420) is a fluorescent cationic dye used to label mitochondria in living cells. Rhodamine 123 inhibits ADP-stimulated respiration of mitochondria with Ki = 12 μM and ATPase activity of inverted inner membrane vesicles with Ki of 126 μM and partially purified F1-ATPase with Ki of 177 μM.

Rhodamine 123 Chemical Structure

Rhodamine 123 Chemical Structure

CAS: 62669-70-9

Selleck's Rhodamine 123 has been cited by 1 publication

Purity & Quality Control

Batch: S357701 DMSO] 76 mg/mL] false] Ethanol] 15 mg/mL] false] Water] Insoluble] false Purity: 99.79%
99.79

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Biological Activity

Description Rhodamine 123 (RH-123, R-22420) is a fluorescent cationic dye used to label mitochondria in living cells. Rhodamine 123 inhibits ADP-stimulated respiration of mitochondria with Ki = 12 μM and ATPase activity of inverted inner membrane vesicles with Ki of 126 μM and partially purified F1-ATPase with Ki of 177 μM.
Targets
ATPase [1]
(inverted inner membrane vesicles)		 F1-ATPase [1]
(Cell-free assay)
126 μM(Ki) 177 μM(Ki)
In vitro
In vitro

1.Preparation of Rhodamine 123 working solution
1.1Preparation of the stock solution
Dissolve 1 mg Rhodamine 123 in 525 μL DMSO to obtain 5 mM of stock solution.
1.2Preparation of Rhodamine 123 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 Rhodamine 123 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.
Cell Research Cell lines KBV200 and HCT-8/V cells with or without knockout of ABCB1
Concentrations 10 μM
Incubation Time 2 h
Method

The cell was seeded into a 6-well plate at a density of 2.5×105 cells/well. After adhered, the cells were then incubated with 10 μM rhodamine 123 or doxorubicin for 2 h at 37°C. After washing three times with ice-cold PBS, cells were analyzed with FCM.

Chemical lnformation & Solubility

Molecular Weight 380.82 Formula
C21H17ClN2O3
CAS No. 62669-70-9 SDF --
Smiles Cl.COC(=O)C1=CC=CC=C1C2=C3C=CC(=N)C=C3OC4=C2C=CC(=C4)N
Storage (From the date of receipt) 3 years -20°C powder

In vitro
Batch:

DMSO : 76 mg/mL ( (199.56 mM)； Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 15 mg/mL

Water : Insoluble


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In vivo
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Working concentration: mg/ml;

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.

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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