610CP TFA

610CP TFA is a new type of actin labeling dye. It dissolves in organic solvents. In DMSO the 610CP TFA excitation/emission wavelength is between 609 and 634 nm. 610CP TFA is a fluorescent dye that penetrates living cells. Upon cell entry, 610CP TFA binds to Bromo-des-methyl-Jasplakinolide Therefore, 610CP TFA dye can be used to stain actin fluorescence images with low background and high resolution.

In Vitro, Guide (The following is our recommended protocol. This protocol only provides guidance and should be modified according to your specific needs). 1. Preparation of 610CP TFA working solution 1.1 Preparation of stock solution Prepare a 10 mM 610CP TFA stock solution using DMSO. Note: It is recommended to aliquot the 610CP TFA stock solution and store at -20°C or -80°C, protected from light. 1.2 Preparation of working solution Dilute the stock solution with pre-warmed serum-free cell culture medium or PBS to prepare a 5-10 μM 610CP TFA working solution. Note: Please adjust the concentration of the 610CP TFA working solution according to actual conditions and prepare fresh before use. 2. Cell staining (suspension cells) 2.1 Collect cells by centrifugation, wash twice with PBS for 5 minutes each time. Adjust the cell density to 1×106/mL. 2.2 Add 1 mL 610CP TFA working solution and incubate at room temperature for 30-60 minutes. 2.3 Centrifuge at 400 g for 3-4 minutes and discard the supernatant. 2.4 Wash the cells twice with PBS for 5 minutes each time. 2.5 Resuspend the cells in 1 mL serum-free medium or PBS, then observe using a fluorescence microscope or flow cytometer. 3. Cell staining (adherent cells) 3.1 Culture adherent cells on sterile coverslips. 3.2 Remove the coverslips from the culture medium and aspirate excess medium. 3.3 Add 100 μL staining working solution, gently shake to ensure complete coverage of the cells, and incubate for 30-60 minutes. 3.4 Remove the staining working solution and wash with culture medium 2-3 times for 5 minutes each time, then observe using a fluorescence microscope.

References:
[1]. Vladimir N Belov, et al. Synthesis of Fluorescent Jasplakinolide Analogues for Live-Cell STED Microscopy of Actin. J Org Chem. 2020 Jun 5;85(11):7267-7275.
[2]. Vladimir N Belov, et al. Synthesis of Fluorescent Jasplakinolide Analogues for Live-Cell STED Microscopy of Actin. J Org Chem. 2020 Jun 5;85(11):7267-7275.