SynaptoRedTM C2

This plan only provides a guide, please modify it to meet your specific needs.

1. Solution preparation

(1) Stock solution: Prepare SynaptoRedTM C2 stock solution with DMSO or EtOH at a concentration of 1-5mM.

Note: After unused stock solution is aliquoted, store it at -20°C or -80°C away from light to avoid repeated freezing and thawing.

(2) Working solution: Dilute the stock solution with a suitable buffer (such as serum-free culture medium, HBSS or PBS) to prepare SynaptoRedTM C2 working solution with a concentration of 5-20μM.

Note: The working solution must be prepared and used immediately. The final concentration of the working solution needs to be optimized according to different cell lines and experimental systems. It is recommended to start from the recommended concentration and explore the optimal concentration in a 10-fold range.

2. Suspended cell staining

(1) Suspended cells are centrifuged at 4°C and 1000rpm for 3-5min and the supernatant is discarded. Wash twice with PBS for 5min each time.

(2) Add 1mL of SynaptoRedTM C2 working solution and incubate at room temperature in the dark for 5-30min. The optimal culture time varies for different cells.

(3) After incubation, centrifuge at 1000-1500rpm for 5min, remove the supernatant, and wash 2-3 times with PBS, 5min each time.

(4) Resuspend the cells in pre-warmed serum-free cell culture medium or PBS. Observe by fluorescence microscopy or flow cytometry.

3. Staining of adherent cells

(1) Culture adherent cells on sterile coverslips.

(2) Remove the coverslip from the culture medium, aspirate the excess culture medium, and place the coverslip in a humid environment.

(3) Add 100μL SynaptoRedTM C2 working solution from one corner of the coverslip and gently shake to evenly cover all cells with the dye.

(4) Incubate at room temperature in the dark for 5-30min. The optimal culture time varies for different cells.

(5) Aspirate the dye working solution and wash the coverslip 2-3 times with culture medium. Observe by fluorescence microscopy.

Note: All fluorescent dyes have quenching problems. Please try to avoid light to slow down fluorescence quenching. For your safety and health, please wear a lab coat and disposable gloves when operating.