FITC-Dextran (MW 10000) (solution)

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).

Labeling of cells^[1]:
Applicable to cells undergoing apoptosis (using HeLa cells and human peripheral blood mononuclear cells (PBMCs) as examples. Note that live HeLa and PBMCs are not stained with FITC-Dextran).
1. Suspend apoptotic cells in 100 μL of culture medium and mix with 10 μL of propidium iodide (PI) and 10 μL of FITC-Dextran (MW 10000) in a Q-prep tube. The final concentrations of PI and FITC-Dextran (MW 10000) are 7.5 μM and 1.13 μM, respectively.
2. Incubate the cells at room temperature in the dark for 25 minutes.
3. Take the labeled cells and centrifuge at 500 g for 10 minutes.
4. Take the centrifuged cells, add 1 mL of culture medium to resuspend them, and analyze them by flow cytometry or fluorescence microscopy (PI: Ex=500 nm, Em=600 nm; FITC-Dextran (MW 10000): Ex=495 nm, Em=525 nm).

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Side-cell permeability detection^[4]
1. Add FITC-Dextran (0.1 mg/mL) to the basal culture medium in the transwell chamber.
2. Collect the culture medium from the transwell inserter after 15 minutes.
3. Measure the fluorescence signal (Ex=495 nm, Em=525 nm).
4. Calculate the FITC-Dextran concentration based on the fluorescence intensity.
5. Calculate the permeability.

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