MetalloFluor™ Series
[Labile ferrous ion detecting probe]
570-590 nm：Orange
FeRhoNox-1 is a fluorescent probe for the detection of iron (II). Because of its high specificity, it can be used to detect intracellular Fe2+. This probe tends to be localized localized in Golgi, and reacts selectively with Fe2+.
This probe does not have chelating effect.
Available through Merck KGaA (Darmstadt, Germany) as:
SCT030 BioTracker™ 575 Red Fe2+ Dye
| Code No. | Product Name | Size | Merck CAT No. |
Merck ( Millipore / Sigma Aldrich ) Product Name |
|---|---|---|---|---|
| GC901 | FeRhoNox™-1 | 50 μg × 10 | SCT030 | BioTracker 575 Red Fe2+ Dye |
FeRhoNox™-1
Product Information
| Product | Target | Cell permeability | reactivity | Absmax (nm) | FLmax (nm) |
| FeRhoNox-1 | Labile irons (Fe2+） | Yes | Irreversible | 540 | 575 |
Figure 1. Response of FeRhoNox-1 to iron(Ⅱ）ion.
Reacting FeRhoNox-1 with Fe2+ for 1 h at 37℃, peak of the fluorescence appears around 575 nm.
Figure 2. The selectivity of FeRhoNox-1.
FeRhoNox-1 can react with only Fe2+ selectively.
FeRhoNox™-1
Cell imaging example using FeRhoNox-1
Live cell imaging of HepG2 cells using FeRhoNox-1 (1) -Fe(II): cells without addition of ferrous ions (100μM ferrous ammonium sulfate) (2) +Fe(II): cells with loaded ferrous ions (3) +Fe(II)+Bpy: further, iron chelator, Bpy was added.
Fluorescence increase was observed in (2), whereas it was decreased in (3). This results is consistent with that FeRhoNox-1 specifically detects Fe(II).
Live cell imaging of HEK293 cells using FeRhoNox-1 (1) -Fe(II): cells without addition of ferrous ions (100μM ferrous ammonium sulfate) (2) +Fe(II): cells with loaded ferrous ions (3) +Fe(II)+Bpy: further, iron chelator, Bpy was added.
Detection of intrinsic ferrous ions. Since fluorescent signal was decreased in (2) Bpy addition, fluorescent signal in (1) represents intrinsic physiological level of ferrous ions.
*Bpy: 2,20-bipyridyl (Bpy) is a cell permeable ferrous ion chelator.
You cannot use for paraffin slices.
To detect Fe2+ ions in tissues, you may prepare frozen section with quick fixation with formaldehyde. Refer the following paper:
Mukaide et al., (2014) Histological detection of catalytic ferrous iron with the selective turn-on fluorescent probe RhoNox-1 in a Fenton reaction-based rat renal carcinogenesis model
Free Radical Research 48:990-995
http://dx.doi.org/10.3109/10715762.2014.898844
We recommend to use dimethyl sulfoxide (DMSO) as the solvent for FeRhoNox-1. Among DMSO, we strongly recommend to use “infinity pure” or “ultra-pure”grade and use freshly opened solvent. You may store aliquots of DMSO at -80 oC and avoid adsorption of humid. Avoid to use with dgradated DMSO, which can increase the background signal of FeRhoNox-1.
The probes are known to be localized either in Golgi or in ER, however, it has been considered that they also detect Fe2+ in the cytoplasmic pool. Please note that the definitive evaluation for this point has not been reported, yet.
Please also refer the frequently asked questions on fluorescent probes
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