PKH 26 

PKH 26 is a red fluorescent dye, PKH 26 can stably bind to the lipid region of cell membrane and emit red fluorescence (Ex/Em=551/567 nm), which is mainly used for in vitro cell labeling, in vitro cell proliferation studies and in vivo and in vitro cell tracing studies^[1].

Guide (The following is the experimental plan we recommend. This plan serves only as a reference guide. The specific operations should be adjusted according to your actual needs.)
1. Preparation of PKH 26 working solution
1.1 Preparation of stock solution
Prepare a 1 mM stock solution using DMSO
1.2 Preparation of PKH 26 working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain a working solution of 5-10 μM.
Note: Please adjust the concentration of the PKH 26 working solution as needed
2. Cell staining
2.1 Suspended cells (6-well plate)
a. Centrifuge at 4°C for 1000g for 3-5 minutes, discard the supernatant, wash twice with PBS, each time for 5 minutes. The cell density is 1×10⁶/mL.
b. Add 1 mL of the working solution, incubate at room temperature for 10-45 minutes.
c. Centrifuge at 400g for 4-5 minutes at 4°C, discard the supernatant.
d. Wash twice with PBS, each time for 5 minutes.
e. Resuspend the cells in serum-free cell culture medium or PBS. Observe with a fluorescence microscope or flow cytometer.
2.2 Attached cells
a. Cultivate attached cells on a sterile cover glass.
b. Remove the cover glass from the culture medium and aspirate the excess medium.
c. Add 100 μL of the working solution, gently shake to fully cover the cells, incubate at room temperature for 5-30 minutes.
d. Wash twice with the culture medium, each time for 5 minutes. Observe with a fluorescence microscope or flow cytometer.

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

961.32

C₅₉H₉₇IN₂

567

551

154214-55-8

Solid

Brown to reddish brown

CC1(C)C(/C=C/C=C2N(CCCCCCCCCCCCCC)C3=C(C=CC=C3)C/2(C)C)=[N+](CCCCCCCCCCCCCCCCCCCCCC)C4=C1C=CC=C4.[I-]

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light, stored under nitrogen

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Fischer K, et al. The flow cytometric PKH-26 assay for the determination of T-cell mediated cytotoxic activity. Methods. 2003;31(2):135-142.  [Content Brief]