Rhod-2 AM

Cat. No.: HY-D0989 Purity: 96.30%: FAQs
Data Sheet SDS COA Handling Instructions

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Rhod-2 is a high-affinity visible light excitation wavelength Ca²⁺ fluorescent probe, Rhod-2, AM is an acetyl methyl ester derivative of Rhod-2, which has cell membrane permeability and can easily enter cells with simple culture. Once it enters the cell, it is sheared by its lactesterase to produce Rhod-2 without membrane permeability, which remains in the cell to perform the corresponding physiological functions. Maximum excitation/emission wavelength: 549/578 nm.

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Rhod-2 AM Chemical Structure

CAS No. : 145037-81-6

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50 μg	USD 60	In-stock	Estimated Time of Arrival: December 31
1 mg	USD 580	In-stock	Estimated Time of Arrival: December 31
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Based on 16 publication(s) in Google Scholar

15 Publications Citing Use of MCE Rhod-2 AM

: Rhod-2 AM purchased from MedChemExpress. Usage Cited in: Environ Sci Technol. 2017 Dec 5;51(23):13938-13948. [Abstract]; Cellular staining with Fluo-3/AM, Rhod-2 and DAPI (blue) in cells upon nLa₂O₃ treatment at 20 μg/mL for 6 h. Cells are examined with confocal microscopy. Two colors are showed individually or merged together.

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Description	Rhod-2 is a high-affinity visible light excitation wavelength Ca²⁺ fluorescent probe, Rhod-2, AM is an acetyl methyl ester derivative of Rhod-2, which has cell membrane permeability and can easily enter cells with simple culture. Once it enters the cell, it is sheared by its lactesterase to produce Rhod-2 without membrane permeability, which remains in the cell to perform the corresponding physiological functions. Maximum excitation/emission wavelength: 549/578 nm^[1].
In Vitro	1.Preparation of Rhod-2 AM working solution 1.1Preparation of the stock solution Dissolve Rhod-2 AM in DMSO to obtain 5 mM of stock solution. 1.2Preparation of Rhod-2 AM working solution Dilute the stock solution in serum-free cell culture medium or PBS to obtain 5-10 μM of working solution. Note: Please adjust the concentration of Rhod-2 AM working solution according to the actual situation. 2.Cell staining (6-well plate) 2.1Suspension cells 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 5-30 minutes. d.Wash twice with medium, 5 minutes each time.Observation by fluorescence microscopy or flow cytometry. MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Rhod-2 AM Related Antibodies
Molecular Weight	1123.94
Formula	C₅₂H₅₉BrN₄O₁₉
CAS No.	145037-81-6
Appearance	Oil
Color	Dark green to black
Emission (Em)	578
Excitation (Ex)	549
SMILES	CN(C1=CC2=[O+]C3=C(C=CC(N(C)C)=C3)C(C4=CC=C(N(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)C(OCCOC5=CC(C)=CC=C5N(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)=C4)=C2C=C1)C.[Br-]
Shipping	Room temperature in continental US; may vary elsewhere.
Storage	-20°C, sealed storage, away from moisture and light *In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)
Purity & Documentation	Purity: ≥97.0% Select Batch: Data Sheet (280 KB) SDS (252 KB) COA (259 KB) Handling Instructions (2659 KB)
Dyeing Example	Description: Rhod-2 AM is a mitochondrial probe, it can be used to detect mitochondrial Ca²⁺ levels with red fluorescence. Method: For mitochondrial potential assessment. 1. After treatment, cells are incubated at 37°C in dark with 4 µM Rhod-2 AM for 20 min. 2. Wash cells for three times and re-suspended in Hank’s Balanced Salt Solution without Ca²⁺ and Mg²⁺. 3. Treat cells with REE NPs and additionally stained with 20 nM TMRM, then analysed by flow cytometry (FACS) for mitochondrial membrane assessment. FACS analysis is performed using an LSR II flow cytometer (Becton Dickinson, Mountain View, CA) equipped with a single 488 nm argon laser. Rhod-2 fluorescence are examined using the PE channel; and both forward and side scatters are used to eliminate cellular fragments. 4. Following treatment, a portion of cells are also fixed with 10% PBS-buffered formaldehyde with 0.1% Triton X-100 for 10 min, then stained with 4’ 6-diamidino-2-phenylindole (DAPI) (blue) and Fluo-3/AM (green) or Rhod-2 (red). Fluorescent images are visualized through confocal laser scanning microscopy. Description: Rhod-2 AM is a mitochondrial probe, it can be used to detect mitochondrial Ca²⁺ levels with red fluorescence. Method: For measurement of mitochondrial Ca²⁺ levels. 1. Cells are incubated with Rhod-2 AM (10 μM; 2 h; dark). 2. Wash cells twice with PBS, and then resuspended in PBS. 3. Fluorescence is determined by flow cytometry, and fluorescence photomicrographs are taken using a fluorescence microscope. Description: Rhod-2 AM is a Ca²⁺ fluorescence probe that specifically aggregates in mitochondria because of its positive charge, it can be used to track mitochondrial Ca²⁺ uptake in cells. Method: For measurement of mitochondrial Ca²⁺ uptake in oocytes. 1. Around 40 oocytes are incubated with Rhod-2 AM (6 µM) in the KSOM medium for 30 min at 37°C. 2. After three quick washes with the KSOM medium, the oocytes are imaged using a Nikon fluorescence microscope (Nikon, Tokyo, Japan).
References	[1]. Brisac C, et al. Calcium flux between the endoplasmic reticulum and mitochondrion contributes to poliovirus-induced apoptosis. J Virol. 2010 Dec;84(23):12226-35. [Content Brief] [2]. Brisac C, et al. Calcium flux between the endoplasmic reticulum and mitochondrion contributes to poliovirus-induced apoptosis. J Virol. 2010 Dec;84(23):12226-35. [Content Brief]

Cell Assay
^[1]

For flow cytofluorometry, cells are harvested, pelleted, and resuspended in ice-cold PBS containing 10 mM glucose, 10% fetal bovine serum (FBS), and 10 μM Rhod-2 AM (Rhod2-AM). Mitochondrial calcium levels are determined by the flow cytofluorometry analysis of aliquots of 4×10⁵ cells. For fluorescence microscopy, IMR5 cells are grown on polylysine-coated (10 μg/mL) slides and stained with 7.5 μM Rhod-2 AM in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS for 2 h before poliovirus (PV) infection. Cells are fixed by incubation for 15 min at 4°C in 4% paraformaldehyde. Cells are washed in PBS, and images are acquired with Zeiss Apotome and Axiovision software^[1].

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

References

[1]. Brisac C, et al. Calcium flux between the endoplasmic reticulum and mitochondrion contributes to poliovirus-induced apoptosis. J Virol. 2010 Dec;84(23):12226-35. [Content Brief]

[2]. Brisac C, et al. Calcium flux between the endoplasmic reticulum and mitochondrion contributes to poliovirus-induced apoptosis. J Virol. 2010 Dec;84(23):12226-35. [Content Brief]

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Rhod-2 AM

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Rhod-2 AM Related Antibodies

15 Publications Citing Use of MCE Rhod-2 AM

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