Cyanine5 DBCO hexafluorophosphate

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)^[1][2].
1. Solution preparation:
1.1 Preparation of storage solution
Solvent: DMSO.
Concentration: 20 mM is recommended.
1.2 Preparation of working solution
Dilute to 20 μM with PBS or serum-free medium (optimized according to the experiment).
(Note: The working solution should be prepared and used immediately, and should be protected from light.)
2. Cell labeling (A549 cells for example):
2.1. Seed cells onto 35-mm glass-bottom dishes at a density of 3×10⁴ (varies by cell) in 2 mL of growth media.
2.2. Add Ac4ManNAz (50 μM, final concentration), incubate for 3 days (generate azide groups on the surface of cells).
2.3. Wash cells twice with DPBS (pH 7.4).
2.4. Incubate cells with Cyanine5 DBCO (20 μM, final concentration) for 1 h at 37°C.
2.5. Rinse cells with DPBS (pH 7.4) and fix with formaldehyde-glutaraldehyde combined fixative for 15 min at 25°C.
2.6. Wash cells twice with DPBS (pH 7.4) and stain cells with DAPI to label nuclei.
2.7. Measure Cyanine5 DBCO fluorescence (Ex=635 nm, Em=650-700 nm) using a confocal laser scanning microscope.
Note:
1) Artificially introduced azide groups on the cell surface can be targeted by Cyanine5 DBCO for more than 3 days after treatment with Ac4ManNAz.
2) Degree of Cyanine5 DBCO labeling is in a dose-dependent manner.

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