Rhod-2, AM *CAS#: 145037-81-6*
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Dissociation constant (Kd, nM) | 570 |
Molecular weight | 1123.96 |
Solvent | DMSO |
Excitation (nm) | 553 |
Emission (nm) | 577 |
Quantum yield | 0.11 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Freeze (< -15 °C); Minimize light exposure |
UNSPSC | 12352200 |
Rhod-2, AM *UltraPure Grade* *CAS#: 145037-81-6* |
Overview | ![]() ![]() |
CAS 145037-81-6 | Molecular weight 1123.96 | Dissociation constant (Kd, nM) 570 | Excitation (nm) 553 | Emission (nm) 577 | Quantum yield 0.11 |
Platform
Fluorescence microscope
Excitation | TRITC filter set |
Emission | TRITC filter set |
Recommended plate | Black wall/clear bottom |
Fluorescence microplate reader
Excitation | 540 |
Emission | 590 |
Cutoff | 570 |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | Bottom read mode/Programmable liquid handling |
Example protocol
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
Prepare a 2 to 5 mM stock solution of Rhod-2 AM in high-quality, anhydrous DMSO.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Rhod-2 AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Rhod-2 AM working solution in a buffer of your choice (e.g., Hanks and Hepes buffer) with 0.04% Pluronic® F-127. For most cell lines, Rhod-2 AM at a final concentration of 4-5 μM is recommended. The exact concentration of indicators required for cell loading must be determined empirically.
Note: The nonionic detergent Pluronic® F-127 is sometimes used to increase the aqueous solubility of Rhod-2 AM. A variety of Pluronic® F-127 solutions can be purchased from AAT Bioquest.
Note: If your cells contain organic anion-transporters, probenecid (1-2 mM) may be added to the dye working solution (final in well concentration will be 0.5-1 mM) to reduce leakage of the de-esterified indicators. A variety of ReadiUse™ Probenecid products, including water-soluble, sodium salt, and stabilized solutions, can be purchased from AAT Bioquest.
SAMPLE EXPERIMENTAL PROTOCOL
Following is our recommended protocol for loading AM esters into live cells. This protocol only provides a guideline and should be modified according to your specific needs.
- Prepare cells in growth medium overnight.
On the next day, add 1X Rhod-2 AM working solution to your cell plate.
Note: If your compound(s) interfere with the serum, replace the growth medium with fresh HHBS buffer before dye-loading.
Incubate the dye-loaded plate in a cell incubator at 37 °C for 30 to 60 minutes.
Note: Incubating the dye for longer than 1 hour can improve signal intensities in certain cell lines.
- Replace the dye working solution with HHBS or buffer of your choice (containing an anion transporter inhibitor, such as 1 mM probenecid, if applicable) to remove any excess probes.
- Add the stimulant as desired and simultaneously measure fluorescence using either a fluorescence microscope equipped with a TRITC filter set or a fluorescence plate reader containing a programmable liquid handling system such as an FDSS, FLIPR, or FlexStation, at Ex/Em = 540/590 nm cutoff 570 nm.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 88.971 µL | 444.856 µL | 889.711 µL | 4.449 mL | 8.897 mL |
5 mM | 17.794 µL | 88.971 µL | 177.942 µL | 889.711 µL | 1.779 mL |
10 mM | 8.897 µL | 44.486 µL | 88.971 µL | 444.856 µL | 889.711 µL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum

Spectral properties
Excitation (nm) | 553 |
Emission (nm) | 577 |
Quantum yield | 0.11 |
Product Family
Name | Excitation (nm) | Emission (nm) |
Fura-2, AM *CAS 108964-32-5* | 336 | 505 |
Fura-2, AM *UltraPure Grade* *CAS 108964-32-5* | 336 | 505 |
Rhod-5N, AM | 557 | 580 |
Rhod-FF, AM | 553 | 577 |
Rhod-4™, AM | 523 | 551 |
Images

Intracellular free Ca2+ was detected by red fluorescent probe dihydrorhod-2 AM (Rhod-2 AM). The nuclei were stained with DAPI (blue). The pEZ-LV203 vector harboring the eGFP reporter gene produced green fluorescent protein. Source: Csseverin inhibits apoptosis through mitochondria-mediated pathways triggered by Ca2+ dyshomeostasis in hepatocarcinoma PLC cells by Shi M et al., PLOS, Nov. 2017.
Citations
Authors: Swift, Luther M and Roberts, Anysja and Pressman, Jenna and Guerrelli, Devon and Allen, Samuel and Haq, Kazi T and Reisz, Julie A and D’Alessandro, Angelo and Posnack, Nikki Gillum
Journal: bioRxiv (2023): 2023--05
Authors: Yoon, Jin-Young and Daneshgar, Nastaran and Chu, Yi and Chen, Biyi and Hefti, Marco and Irani, Kaikobad and Song, Long-Sheng and Brenner, Charles and Abel, E Dale and London, Barry and others,
Journal: bioRxiv (2022)
Authors: Yang, Yuting and Wu, Jihong and Lu, Wei and Dai, Yiqin and Zhang, Youjia and Sun, Xinghuai
Journal: (2022)
Authors: Miranda, Jose G and Schleicher, Wolfgang E and Wells, Kristen L and Ramirez, David G and Landgrave, Samantha P and Benninger, Richard KP
Journal: Molecular metabolism (2022): 101430
Authors: Miranda, Jose G and Schleicher, Wolfgang E and Ramirez, David G and Landgrave, Samantha P and Benninger, Richard KP
Journal: BioRXiv (2020)
Authors: G{\'o}mez-Cid, Lidia and Moro-L{\'o}pez, Marina and de la Nava, Ana S and Hern{\'a}ndez-Romero, Ismael and Fern{\'a}ndez, Ana I and Su{\'a}rez-Sancho, Susana and Atienza, Felipe and Grigorian-Shamagian, Lilian and Fern{\'a}ndez-Avil{\'e}s, Francisco
Journal: Processes (2020): 924
Authors: Wang, Ruifang and Zhu, Yinxing and Lin, Xian and Ren, Chenwei and Zhao, Jiachang and Wang, Fangfang and Gao, Xiaochen and Xiao, Rong and Zhao, Lianzhong and Chen, Huanchun and others,
Journal: Autophagy (2019): 1163--1181
Authors: Swift, Luther M and Jaimes III, Rafael and McCullough, Damon and Burke, Morgan and Reilly, Marissa and Maeda, Takuya and Zhang, Hanyu and Ishibashi, Nobuyuki and Rogers, Jack M and Posnack, Nikki Gillum
Journal: Journal of visualized experiments: JoVE (2019)
Authors: Birkner, Antje
Journal: (2019)
Authors: Vierra, Nicholas C and Kirmiz, Michael and van der List, Deborah and Santana, L Fernando and Trimmer, James S
Journal: Elife (2019): e49953
References
Authors: Stamm C, del Nido PJ.
Journal: Thorac Cardiovasc Surg (2004): 127
Authors: Stamm C, Friehs I, Choi YH, Zurakowski D, McGowan FX, del Nido PJ.
Journal: Cardiovasc Res (2003): 695
Authors: Du C, MacGowan GA, Farkas DL, Koretsky AP.
Journal: Biophys J (2001): 549
Authors: Du C, MacGowan GA, Farkas DL, Koretsky AP.
Journal: Cell Calcium (2001): 217
Authors: Lannergren J, Westerblad H, Bruton JD.
Journal: J Muscle Res Cell Motil (2001): 265
Authors: MacGowan GA, Du C, Glonty V, Suhan JP, Koretsky AP, Farkas DL.
Journal: J Biomed Opt (2001): 23
Authors: Muriel MP, Lambeng N, Darios F, Michel PP, Hirsch EC, Agid Y, Ruberg M.
Journal: J Comp Neurol (2000): 297
Authors: Del Nido PJ, Glynn P, Buenaventura P, Salama G, Koretsky AP.
Journal: Am J Physiol (1998): H728
Application notes
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