Cy3 Dextran (MW 10000)

Guide (The following is our recommended solution. This solution is merely a guideline and should be modified according to your specific needs.)
Preparation of Cy3-Dextran working solution
Dissolve it in sterile PBS or serum-free medium. The recommended working concentration is 5–50 μg/mL.
If sterilization or removal of impurities is required, filter with a 0.22 μm filter membrane. It is recommended to use the prepared solution within 2 hours.
2. Cell Incubation and Washing
2.1 Remove the original culture medium of the cells and gently rinse the cells with sterile PBS 2-3 times to remove any residual serum (serum proteins can interfere with the background).
2.2 Add the prepared Cy3-Dextran working solution and ensure that it completely covers the cell layer.
2.3 The samples were incubated in a 37°C, 5% CO₂ incubator under dark conditions for 2 to 4 hours (the exact duration was adjusted according to the type of cells: 2-3 hours for tumor cells, and could be slightly longer for normal cells).
2.4 After incubation is completed, discard the working solution. Gently rinse the cells 3-4 times with sterile PBS, each time for 1-2 minutes, to thoroughly remove the unabsorbed free dyes and reduce background fluorescence.
2.5 (Optional) If you need to perform fixed preservation, add 4% formaldehyde to fix for 15-20 minutes, then wash with PBS twice.
3. Fluorescence Observation and Collection
3.1 Place the culture dish under the fluorescence microscope and select the Cy3 corresponding channel (Ex/Em = 550/570 nm).
3.2 Appropriately adjust the exposure time to prevent overexposure from causing photobleaching.
3.3 Collect the orange-red fluorescence signals within the cells. This can be combined with software such as ImageJ for quantitative analysis of fluorescence intensity to evaluate the cell uptake efficiency.

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