2-NBDG (solution)

Guide (The following is the experimental plan we recommend. This plan serves only as a reference guide. The specific operations should be adjusted according to your actual needs.)
1. Preparation of 2-NBDG Solution Dilute the stored solution with pre-warmed serum-free cell culture medium or PBS to prepare a 2-NBDG working solution of 10-200 μM.
Note: Please adjust the concentration of the 2-NBDG working solution according to the actual situation, and prepare it as needed.
2. Cell Staining 2.1 Preparation of Cells Suspended cells: Centrifuge at 1000 g for 3-5 minutes at 4°C. Collect the cell precipitate and wash it twice with PBS, each time for 5 minutes.
Monolayer cells: Remove the culture medium and add trypsin to digest the cells. Centrifuge at 1000 g for 3-5 minutes at 4°C. Collect the cell pellet, add PBS for two washes, each for 5 minutes.
2.2 Add 1 mL of the 2-NBDG working solution and incubate at room temperature for 5 to 60 minutes.
2.3 400 g, centrifuge at 4℃ for 3-4 minutes, then discard the supernatant.
2.4 Add PBS to wash the cells twice, for 5 minutes each time.
2.5 After resuspending the cells in 1 mL of serum-free medium or PBS, observe them using a flow cytometer;
If an activity test is conducted, use an ELISA microplate reader to record the optical density (O.D.) at 540/570 nm. The cell viability is calculated based on the control ratio, and a curve is plotted against the logarithmic concentration of the drug. The IC₅₀ value is then calculated.

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