Rhod-4™, potassium salt

ATP-stimulated calcium responses of endogenous P2Y receptors were measured in CHO-K1 cells with Rhod-4™ AM (Cat# 21120) and Rhod-2 AM (Cat# 21064). CHO-K1 cells were seeded overnight at 50,000 cells/100 µL/well in a Costar 96-well black wall/clear bottom plate. The growth medium was removed, and the cells were incubated with 100 µL of dye loading solution using Rhod-4™ AM (4 µM, A and B) or Rhod-2 AM (4 µM, C and D) for 1 hour in a 37 °C, 5% CO2 incubator. The staining solution was replaced with 200 µL HHBS, then the cells were imaged before (A and C) and after (B and D) ATP treatment with a fluorescence microscope (Olympus IX71) using TRITC channel.

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Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
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Physical properties

Dissociation constant (K_d, nM)	451
Molecular weight	880.07
Solvent	Water

Spectral properties

Excitation (nm)	523
Emission (nm)	551
Quantum yield	0.1¹

Storage, safety and handling

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

Overview SDS Protocol

Molecular weight

880.07

Dissociation constant (K_d, nM)

451

Excitation (nm)

523

Emission (nm)

551

Quantum yield

0.1¹

Calcium measurement is critical for numerous biological investigations. Fluorescent probes that show spectral responses upon binding Ca2+ have enabled researchers to investigate changes in intracellular free Ca2+ concentrations by using fluorescence microscopy, flow cytometry, fluorescence spectroscopy and fluorescence microplate readers. Rhod-2 is most commonly used among the red fluorescent calcium indicators. However, Rhod-2 AM is only moderately fluorescent in live cells upon esterase hydrolysis, and has very small cellular calcium responses. Rhod-4™ has been developed to improve Rhod-2 cell loading and calcium response while maintaining the spectral wavelength of Rhod-2. In CHO and HEK cells Rhod-4™ AM has cellular calcium response that is 10 times more sensitive than Rhod-2 AM. AAT Bioquest offers versatile packing sizes of Quest Rhod-4 to meet your special needs, e.g., 1 mg; 10x50 µg; 20x50 µg; HTS packages with no additional packaging charge.

Calculators

Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of Rhod-4™, potassium salt to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	113.627 µL	568.137 µL	1.136 mL	5.681 mL	11.363 mL
5 mM	22.725 µL	113.627 µL	227.255 µL	1.136 mL	2.273 mL
10 mM	11.363 µL	56.814 µL	113.627 µL	568.137 µL	1.136 mL

Molarity calculator

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Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
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Spectrum

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Spectral properties

Excitation (nm)	523
Emission (nm)	551
Quantum yield	0.1¹

Product Family

Name	Excitation (nm)	Emission (nm)	Quantum yield
Rhod-4™, sodium salt	523	551	0.1¹

Images

Figure 1. ATP-stimulated calcium responses of endogenous P2Y receptors were measured in CHO-K1 cells with Rhod-4™ AM (Cat# 21120) and Rhod-2 AM (Cat# 21064). CHO-K1 cells were seeded overnight at 50,000 cells/100 µL/well in a Costar 96-well black wall/clear bottom plate. The growth medium was removed, and the cells were incubated with 100 µL of dye loading solution using Rhod-4™ AM (4 µM, A and B) or Rhod-2 AM (4 µM, C and D) for 1 hour in a 37 °C, 5% CO2 incubator. The staining solution was replaced with 200 µL HHBS, then the cells were imaged before (A and C) and after (B and D) ATP treatment with a fluorescence microscope (Olympus IX71) using TRITC channel.

Citations

View all 23 citations: Citation Explorer

Effect of stem cell niche elasticity/ECM protein on the self-beating cardiomyocyte differentiation of inducedpluripotent stem (iPS) cells at different stages
Authors: Hirata, Mitsuhi and Yamaoka, Tetsuji
Journal: Acta Biomaterialia (2017)

Emerin plays a crucial role in nuclear invagination and in the nuclear calcium transient
Authors: Shimojima, Masaya and Yuasa, Shinsuke and Motoda, Chikaaki and Yozu, Gakuto and Nagai, Toshihiro and Ito, Shogo and Lachmann, Mark and Kashimura, Shin and Takei, Makoto and Kusumoto, Dai and others, undefined
Journal: Scientific Reports (2017)

Preliminary findings on ultrasound modulation of the electromechanical function of human stem-cell-derived cardiomyocytes
Authors: Chen, Andrew William and Klimas, Aleks and ra , undefined and Zderic, Vesna and Castellanos, Ivan Suares and Entcheva, Emilia
Journal: (2017): 1--4

The role of spatial organization of Ca (2+) release sites in the generation of arrhythmogenic diastolic Ca (2+) release in myocytes from failing hearts.
Authors: Belevych, Andriy E and Ho, Hsiang-Ting and Bonilla, Ingrid M and Terentyeva, Radmila and Schober, Karsten E and Terentyev, Dmitry and Carnes, Cynthia A and Györke, Sándor
Journal: Basic research in cardiology (2017): 44

Individual evaluation of cardiac marker expression and self-beating during cardiac differentiation of P19CL6 cells on different culture substrates
Authors: Yamaoka, Tetsuji and Hirata, Mitsuhi and Dan, Takaaki and Yamashita, Atsushi and Otaka, Akihisa and Nakaoki, Takahiko and Miskon, Azizi and Kakinoki, Sachiro and Mahara, Atsushi
Journal: Journal of Biomedical Materials Research Part A (2016)

Multiple pathways for elevating extracellular adenosine in the rat hippocampal CA1 region characterized by adenosine sensor cells
Authors: Yamashiro, Kunihiko and Fujii, Yuki and Maekawa, Shohei and Morita, Mitsuhiro
Journal: Journal of Neurochemistry (2016)

The G protein-coupled receptor GPR157 regulates neuronal differentiation of radial glial progenitors through the Gq-IP3 pathway
Authors: Takeo, Yutaka and Kurabayashi, Nobuhiro and Nguyen, Minh Dang and Sanada, Kamon
Journal: Scientific reports (2016)

Dynamic polyrotaxane-coated surface for effective differentiation of mouse induced pluripotent stem cells into cardiomyocytes
Authors: Seo, Ji-Hun and Hirata, Mitsuhi and Kakinoki, Sachiro and Yamaoka, Tetsuji and Yui, Nobuhiko
Journal: RSC Advances (2016): 35668--35676

Involvement of aberrant calcium signalling in herpetic neuralgia
Authors: Warwick, Rebekah A and Hanani, Menachem
Journal: Experimental neurology (2016): 10--18

OptoDyCE as an automated system for high-throughput all-optical dynamic cardiac electrophysiology
Authors: Klimas, Aleks and ra , undefined and Ambrosi, Christina M and Yu, Jinzhu and Williams, John C and Bien, Harold and Entcheva, Emilia
Journal: Nature communications (2016)