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Cyanine5 DBCO (DBCO-Cy5) is a low-toxicity azide reactive probe (NIR fluorescent dye), for imaging azide-labeled biomolecules via a copper-free "click-through" reaction. Cyanine5 DBCO has no apparent cytotoxicity or animal toxicity and shows no damage to the physiological functions of cells other than the target cells (azide-labeled cells). Cyanine5 DBCO can be used to label and track cells in vitro and in vivo (Ex=635 nm, Em=650-700 nm)[1][2]. | [in vivo]
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
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Cell tracking in vivo[1][2]:
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1. Seed cells onto 35-mm glass-bottom dishes at a density of 3×104 (varies by cell) in 2 mL of growth media.
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2. Add Ac4ManNAz (50 μM, final concentration), incubate for 3 days (generate azide groups on the surface of cells).
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3. Wash cells twice with DPBS (pH 7.4).
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4. Anesthesia for mice, then inject cells (after Ac4ManNAz treatment) into the left lobe of the liver.
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5. Inject Cyanine5 DBCO (25 μM, 200 μL) to tail vein (3 days post injection of cells into the liver).
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6. Measure Cyanine5 DBCO fluorescence (Ex=635 nm, Em=650-700 nm). In vivo near-infrared fluorescence (NIRF) images is obtained with the in vivo imaging system. | [References]
[1] Kang SW, et al. Cell labeling and tracking method without distorted signals by phagocytosis of macrophages. Theranostics. 2014 Feb 12;4(4):420-31. DOI:10.7150/thno.7265
[2] Zhu L, et al. Coupling Aptamer-based Protein Tagging with Metabolic Glycan Labeling for In Situ Visualization and Biological Function Study of Exosomal Protein-Specific Glycosylation. Angew Chem Int Ed Engl. 2021 Aug 9;60(33):18111-18115. DOI:10.1002/anie.202103696
[3] Song S, et al. In Situ One-Step Fluorescence Labeling Strategy of Exosomes via Bioorthogonal Click Chemistry for Real-Time Exosome Tracking In Vitro and In Vivo. Bioconjug Chem. 2020 May 20;31(5):1562-1574. DOI:10.1021/acs.bioconjchem.0c00216 |
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