PKH67 Membrane Dye Labeling Kit (#PKH67GL, Sigma‐Aldrich) was used to label sEVs with green fluorescence to track the localization of sEVs. Briefly, sEVs were resuspended in provided diluent C. PKH67 reagent was added to diluent C prior to mixing with sEVs. The mixtures were supplied with 1% BSA to eliminate the excess dye, and ultrafiltered using 100 kDa Amicon Ultra‐15 Centrifugal Units (#UFC910024, Merck Millipore) at 3,000 × g for 30 min at 4°C. Subsequently, the samples were resuspended in PBS and harvested by ultracentrifugation.
Pkh67 membrane dye labeling kit
Manufactured by Merck Group
The PKH67 Membrane Dye Labeling Kit is a laboratory product designed for fluorescent labeling of cells. The kit contains a fluorescent dye that binds to the lipid bilayer of cell membranes, allowing for the visualization and tracking of labeled cells.
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Lab products found in correlation
2 protocols using pkh67 membrane dye labeling kit
1
PKH67 Labeling of Small Extracellular Vesicles
2
Quantifying sEV Internalization in HUVECs
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sEVs were labeled with PKH‐67 Membrane Dye Labeling Kit (Sigma–Aldrich) according to manufacturer's protocol with minor modifications. To assess the number of sEVs taken up by HUVECs, 5 × 104 cells were seeded in 6‐well plate with cover slips for 24 h. An amount of 2.5 µg of PKH67‐labeled sEVs in serum‐free medium was added as treatment. After 2 h of incubation, sEV‐treated HUVECs were washed by PBS for five times followed by fixation with 4% paraformaldehyde and staining with phalloidin and DAPI. The coverslips were mounted onto slides and examined under laser scanning confocal microscopy. Images were analyzed using ImageJ. The relative PKH67‐sEVs uptake was determined by mean intensity of fluorescent signal from at least three different fields that were randomly selected from each sample.
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