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36104
FerroOrange
Buffers & Dyes
Cellular Dyes
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FerroOrange #36104

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Confocal analysis of HeLa cells, untreated (left), treated with ammonium iron(II) sulfate (100 μM, 30 min; middle), or inhibited with 2,2'-bipyridine (100 μM) after treating with ammonium iron(II) sulfate (100 μM, 30 min; right) using FerroOrange (orange) and Hoechst 33342 #4082 (blue).
Flow cytometric analysis of live RAW 264.7 cells, stained with FerroOrange (solid line) or unstained (dashed line).
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Product Usage Information

Reagent Preparation:

Material appears as a clear to whitish solid/film that is difficult to see in the tube and can consolidate within the cap. To ensure material has been contained within the vial, centrifuge the tube of lyophilized FerroOrange once it has been brought to room temperature and prior to reconstitution. To create a 1 mmol/L FerroOrange solution, combine 35 µL of DMSO with 24 µg of FerroOrange and pipette repeatedly to dissolve. To prepare the 1 µmol/L solution of FerroOrange, dilute the 1 mmol/L FerroOrange 1:1000 in HBSS. Protect from light.

IMPORTANT: DMF or ethanol can be used as an alternative to DMSO. Serum-free medium can be used as an alternative to HBSS. Serum-containing medium cannot be used as it will generate high background. The 1 mmol/L solution is stable for 1 month at -20°C protected from light. The 1 µmol/L solution is unstable and should be prepared and used immediately before staining the experiment.

NOTE: If working with suspension cells, it is necessary to pellet the cells by gentle centrifugation before all wash and aspiration steps.

Protocol:

1. Culture cells as appropriate, and perform treatments as needed.

2. Aspirate the supernatant.

3. Wash the cells three times with HBSS or serum-free media.

4. Add the 1 µmol/L FerroOrange working solution to the cells and incubate for 30 min.

5. Examine cells by fluorescence microscopy. Refer to the Fluorescent Properties section for excitation and emission spectra.

6. For analysis by flow cytometry, dissociate adherent cells or collect suspension cells. Analyze on a flow cytometer at desired cell density.

Storage

Store at 4°C desiccated and protected from light and metal. In lyophilized form, the product is stable for 12 months. Once reconstituted at a concentration of 1 mmol/L, store at -20ºC protected from light and metal, and use within 1 month.

Fluorescent Properties

  • Excitation maximum = 543 nm/Emission maximum = 580 nm

Product Description

FerroOrange is a small molecular iron sensing dye containing a fluorescent probe that selectively binds iron ions, allowing for detection of intracellular iron via live-cell fluorescent imaging. When the iron ions are bound to FerroOrange, fluorescence increases, resulting in the detection and measurement of intracellular iron. It has been shown that free iron in living cells (mostly Fe2+) may be related to cellular damage or death. Iron exists in ferrous and ferric states, and FerroOrange uniquely detects the labile ferrous form, which drives ferroptosis. Ferroptosis is a distinct type of cell death defined by iron-dependent lipid peroxidation. FerroOrange aids in ferroptosis research and detection.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not for Use in Diagnostic Procedures.
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To Purchase # 36104
Cat. # Size Qty. Price
36104S		 24 µg

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