Phen Green SK diacetate (5/6-mixture)

Phen Green SK diacetate (5/6-mixture) (20 μM; 10 min) chelates Fe²⁺ in rat hepatocytes, resulting in fluorescence quenching. However, fluorescence was restored after the addition of 5 mM 2,2'-Dipyridyl^[1].
The fluorescence of Phen Green SK diacetate (5/6-mixture) (20 μM; 10 min) in human erythroleukemia K562 cells was quenched by 10 μM Fe(III)-8-Hydroxyquinoline (HY-B1005) complex, and the iron concentration in K562 cells was measured to be 4.0 μM^[1].

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)^[2][3].
1. Solution preparation
1.1 Preparation of stock solution
Solvent: DMSO
Concentration: 20 mM (optimized according to the experiment).
1.2 Preparation of working solution
Dilute to 10-20 μM with PBS or cell culture medium.
Note: The working solution should be prepared and used immediately. Keep it away from light.
2. Phen Green SK diacetate (5/6-mixture) is used to detect intracellular Fe²⁺ levels:
2.1 Wash the target cells (e.g., HepG2 and Huh7 cells, H9C2 cells) with phosphate-buffered saline (PBS) and incubate with Phen Green SK diacetate (5/6-mixture) at 37 °C in the dark (the appropriate concentration for HepG2 and Huh7 cells is 20 μM, 20 min; the appropriate concentration for H9C2 cells is 10 μM, 30 min).
2.2 After washing the cells again, suspend the cells in PBS and use them for flow cytometry analysis.

Phen Green SK diacetate (5/6-mixture) can be used in conjunction with the lipid peroxidation indicator dye BODIPY 581/591 C11 (HY-D1301) to assess the ferroptosis state of cells^[3].

Effects of different metal and nonmetal ions on the fluorescence of Phen Green SK diacetate (5/6-mixture) in a cell-free system (spectrofluorimetry)^[1]:
(1) PG SK (50 μM) was dissolved in Chelex-treated medium (pH 7.2, 37°C) designed to mimic the composition of cytosol. (2) Baseline fluorescence was recorded for 5 min (excitation wavelength: 488 nm; emission wavelength: 520 nm). (3) The corresponding salt ions were added and quickly mixed with the medium in the cuvette. The relative emission intensity was expressed as a percentage of the baseline fluorescence intensity (set to 100%).
Results show^[1]:

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.