Biotracker far red labile fe2 dye

Manufactured by Merck Group

BioTracker Far-red Labile Fe2+ dye is a fluorescent probe designed for the detection and monitoring of labile iron(II) ions in biological systems. The dye exhibits a shift in its fluorescence emission spectrum upon binding to ferrous iron, allowing for the visualization and quantification of iron(II) levels.

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Lab products found in correlation

Bodipy c11 reagent, by Thermo Fisher Scientific (1 mentions) Ebioscience intracellular fixation and permeabilization buffer set, by Thermo Fisher Scientific (1 mentions) Cellrox green reagent, by Thermo Fisher Scientific (1 mentions) C11 bodipy, by Thermo Fisher Scientific (1 mentions) Accuri c6 flow cytometer, by BD (1 mentions) Navios flow cytometer, by BD (1 mentions) Cflow plus software, by BD (1 mentions) Bodipy 581 591 c11, by Thermo Fisher Scientific (1 mentions)

2 protocols using biotracker far red labile fe2 dye

Multimodal Characterization of Breast Cancer Cells in Co-Culture with MSCs

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We separated breast cancer cells in co-culture conditions from MSCs using human EpCAM microbeads as described above for CSC and ER staining experiments. We performed CSC staining and analysis as previously described (36 (link)).
We stained for ER-α using [E115]-ChIP Grade (#ab32063, Abcam, Waltham, MA, USA) antibody at the manufacturer’s recommended dilution. We then fixed and permeabilized cells using eBioscience Intracellular Fixation and Permeabilization Buffer Set (#88-8824-00, Invitrogen, Waltham, MA, USA). We based flow cytometry gates on secondary only control.
We measured reactive oxygen species (ROS) using CellROX Green Reagent (#C10444, ThermoFisher), labile iron using BioTracker Far-red Labile Fe2+ dye (#SCT037, Sigma Aldrich), intracellular Zn2+ using BioTracker 515 Green Zn2+ DA Dye (#SCT032, Sigma Aldrich), intracellular Cu+ using BioTracker Green Copper Live Cell Dye (#SCT041, Sigma Aldrich), and relative lipid ROS using BODIPY-C11 reagent (#C10445, ThermoFisher) following manufacturer’s recommendations. We separated signals from cancer cells and stromal cells by GFP-tagged cancer cells for iron, mCherry-NLS-tagged cancer cells for ROS, zinc, and copper stains, and BFP-NLS-tagged cancer cells for lipid ROS.

Buschhaus J.M., Rajendran S., Humphries B.A., Cutter A.C., Muñiz A.J., Ciavattone N.G., Buschhaus A.M., Cañeque T., Nwosu Z.C., Sahoo D., Bevoor A.S., Shah Y.M., Lyssiotis C.A., Ghosh P., Wicha M.S., Rodriguez R, & Luker G.D. (2022). Effects of iron modulation on mesenchymal stem cell-induced drug resistance in estrogen-receptor-positive breast cancer. Oncogene, 41(29), 3705-3718.

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Measuring Iron Levels and Lipid Peroxidation in Cells

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Data acquisition and data analysis were conducted at the Cochin Cytometry and Immunobiology Facility. For iron assay measurements using BioTracker Far-red Labile Fe2+ Dye (called FerroFarRed, #SCT037, Sigma; Thermofischer; Waltham, MA), 2 × 10⁵ cells were washed twice in PBS to remove extracellular Fe2+ then labeled with 2 µM Sirhonox for 60 min in a tissue culture incubator (37 °C, 5% CO²) in the dark and then washed twice again. The pelleted cells were then resuspended in 0.3 mL of PBS. FCM data were collected using a BD Navios flow cytometer. 10,000 events were recorded for analysis. Data analysis was then carried out with KALUZA software. For lipid peroxide production measurements using C11- BODIPY (581/591) (2 mM) (Thermofischer, Waltham, MA), 2 × 10⁵ cells were labeled with C11-BODIPY in 1 mL of warm complete medium for 10 min in a tissue culture incubator (37 °C, 5% CO²) in the dark. Cells were then washed twice and resuspended in 200 µL of fresh PBS. FCM data were collected using a C6 Accuri flow cytometer (Becton Dickinson, Le Pont de Claix, France) with CFlow Plus software. 10,000 events were captured for subsequent analysis with CFlow Plus software (Becton Dickinson, Le Pont de Claix, France).

Grignano E., Cantero-Aguilar L., Tuerdi Z., Chabane T., Vazquez R., Johnson N., Zerbit J., Decroocq J., Birsen R., Fontenay M., Kosmider O., Chapuis N, & Bouscary D. (2023). Dihydroartemisinin-induced ferroptosis in acute myeloid leukemia: links to iron metabolism and metallothionein. Cell Death Discovery, 9, 97.

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