Cyanine5 DBCO (DBCO-Cy5) 是一种水溶性的，低毒的叠氮化物反应探针 (近红外荧光染料)，可通过无铜“点击反应”对叠氮化物标记的生物分子进行成像 (Ex=635 nm, Em=650-700 nm)。Cyanine5 DBCO 无明显的细胞和动物毒性，并且不影响靶细胞 (叠氮化标记细胞) 以外的其他细胞的生理功能。Cyanine5 DBCO 可用于体内外细胞的标记和追踪。

Cyanine5 DBCO (DBCO-Cy5) is a water-soluble, 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)

Solid

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Cell labeling (A549 cells for example):
1. Seed cells onto 35-mm glass-bottom dishes at a density of 3×10 (varies by cell) in 2 mL of growth media.
2. Add Ac4ManNAz (50 μM, final concentration), incubate for 3 days (generate azide groups on the surface of cells).
3. Wash cells twice with DPBS (pH 7.4).
4. Incubate cells with Cyanine5 DBCO (20 μM, final concentration) for 1 h at 37℃.
5. Rinse cells with DPBS (pH 7.4) and fix with formaldehyde-glutaraldehyde combined fixative for 15 min at 25℃.
6. Wash cells twice with DPBS (pH 7.4) and stain cells with DAPI to label nuclei.
7. Measure Cyanine5 DBCO fluorescence (Ex=635 nm, Em=650-700 nm) using a confocal laser scan

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Cell tracking in vivo:
1. Seed cells onto 35-mm glass-bottom dishes at a density of 3×10 (varies by cell) in 2 mL of growth media.
2. Add Ac4ManNAz (50 μM, final concentration), incubate for 3 days (generate azide groups on the surface of cells).
3. Wash cells twice with DPBS (pH 7.4).
4. Anesthesia for mice, then inject cells (after Ac4ManNAz treatment) into the left lobe of the liver.
5. Inject Cyanine5 DBCO (25 μM, 200 μL) to tail vein (3 days post injection of cells into the liver).
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.

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light, stored under nitrogen In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)

[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.
[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.

In Vitro: DMSO : 125 mg/mL (134.55 mM; Need ultrasonic)配制储备液