The 786-O-EVs were labeled by a PKH26 red fluorescence labeling kit (Sigma-Aldrich; Merck KGaA) in compliance with the provided instructions. Briefly, the EVs underwent a 4-min incubation regimen with the PKH26 dye, which was terminated by the addition of EV-free FBS. The EVs were rinsed three times and the excess PKH26 dye was removed with an Amicon Ultra-4 (100-kDa, Merck Millipore Corp.), followed by a 4-h incubation regimen with the RCC cells. The internalization of EVs by RCC cells was observed under confocal microscopy.
Pkh26 red fluorescence labeling kit
Manufactured by Merck Group
Sourced in United States
The PKH26 red fluorescence labeling kit is a reagent used for the fluorescent staining of cells. It provides a stable red fluorescent label that can be detected using flow cytometry or fluorescence microscopy. The kit includes the PKH26 dye and associated solutions for efficient cell labeling.
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Lab products found in correlation
2 protocols using pkh26 red fluorescence labeling kit
1
Fluorescent Labeling of Extracellular Vesicles for Uptake Studies
2
Exosome Labeling and Internalization
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For internalization of exosomes derived from AT-MSCs, EVs were labeled with a PKH26 red fluorescence labeling kit according to the manufacturer’s instruction (Sigma-Aldrich, Thermo Fisher Scientific, Waltham, MA, USA). Briefly, 30 µg of exosomes in 90 µL of PBS were mixed with 360 µL of 8 µM PKH26 and incubated for 5 min with mixing by gentle pipetting. The excess dye was absorbed with 360 µL of 1% BSA in PBS. Then, the EVs were washed with PBS at 110,000× g for 70 min. The pellets were resuspended in 500 µL of PBS and transferred into a 100-kDa filter (Microcon YM-100, Millipore, Merck Sigma, MA, USA), and the cell suspension was concentrated to 30 µL. PKH26-labeled EVs were added to the recipient cells. After 24 h of incubation, images were acquired using confocal microscopy (FV10i; Olympus Life Science, Tokyo, Japan).
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