HKSOX-1r (5/6-mixture)

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of working solution.
1. Preparation of HKSOX-1r (5/6-mixture) working solution
1.1 Preparation of the stock solution
Dissolve 1 mg HKSOX-1r (5/6-mixture) in 117 μL DMSO to obtain 10 mM of stock solution.
Note: It is recommended to store the stock solution at -20°C -80°C away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of HKSOX-1r (5/6-mixture) working solution
Note: The working solution should be prepared and used immediately
2. Cell staining
2.1 Suspension cells (6-well plate)
a. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×10⁶/mL
b. Add 1 mL of working solution, and then incubate at room temperature for 5-30 minutes.
c. Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
d. Wash twice with PBS, 5 minutes each time.
e. Resuspend cells with serum-free cell culture medium or PBS.Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
a. Culture adherent cells on sterile coverslips.
b. Remove the coverslip from the medium and aspirate excess medium.
c. Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 5-30 minutes.
d. Wash twice with medium, 5 minutes each time.Observation by fluorescence microscopy or flow cytometry.


HKSOX-1r 5/6-mixture (2 μM; 30 min) detects mitochondrial respiratory inhibitor-induced O₂^?? formation in a highly sensitive and rapid manner in HCT116, BV-2 and RAW264.7 cells^[1].

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