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].

General Protocol
1.Preparation of PKH 26 working solution
1.1Preparation of the stock solution
Dissolve 1 mg PKH 26 in 1 mL DMSO to obtain 1 mM of stock solution.
Note: It is recommended to store the stock solution at -20°C and -80°C away from light and avoid repetitive freeze-thaw cycles.
1.2Preparation of PKH 26 working solution
Dilute the stock solution in serum-free cell culture medium or PBS dilute at 1:50 or 1:100 to obtain 5-10 μM of working solution.
Note: Please adjust the concentration of PKH 26 working solution according to the actual situation.
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 10-45 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.

Storage
-80°C, 1 year
Protect from light

Precautions
1. Please adjust the concentration of PKH 26 working solution according to the actual situation.
2. This product is for R&D use only, not for drug, household, or other uses.
3. For your safety and health, please wear a lab coat and disposable gloves to operate.

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

961.32

C₅₉H₉₇IN₂

154214-55-8

Solid

Brown to reddish brown

567

551

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]