EC-17 disodium salt

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
1.1 Preparation of storage solution ^[2]
Prepare EC-17 disodium salt stock solution.
Note: EC-17 storage solution is recommended to be stored at -20°C or -80°C in the dark after aliquoting.
1.2 Preparation of working solution
Dilute the storage solution with cell suspension or blood sample to make the final EC-17 disodium salt concentration of 200 nM.
Note: Please adjust the concentration of EC-17 disodium salt working solution according to the actual situation and prepare it before use.
2 Staining
2.1 In vitro cell staining
1) FR⁺ L1210A or KB cells were suspended at a concentration of 10⁶ cells/mL in PBS containing 2% fetal bovine serum (FBS).
2) 1 mL of cell suspension is added with 20 μL of 10 μM EC-17 disodium salt stock solution to a final concentration of 200 nM.
3) The mixture is incubated at 37°C for 60 min.
4) The cells are washed twice with PBS and then resuspended at a concentration of 10⁶ cells/mL for subsequent analysis.
2.2 Whole blood sample staining
1) Blood is drawn from female nude mice and stabilized with 1000 units/mL Heparin (HY-17567) at a 1:1 dilution.
2) 10⁴ circulating tumor cells (CTCs) are added to a 500 μL blood aliquot and the sample is mixed by gentle vortexing for 30 s.
3) 10 μL of 10 μM EC-17 disodium salt stock solution is added to 0.5 mL blood sample for a final concentration of 200 nM.
4) The blood-cell suspension is incubated at 37°C for 60 min.
5) To test labeling specificity, cells are pre-incubated with 10 μM free Folic acid (HY-16637) before adding EC-17.
6) Using a flow cytometer, analyze the labeled cells using the cyan (488 nm laser, 530 nm emission filter) and red (637 nm laser, 647 nm emission filter) channels.

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