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。
| 生物活性 | 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)[1][2]. |
体外研究
(In Vitro) | 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][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. Add Ac4ManNAz (50 μM, final concentration), incubate for3 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) for1 hat 37℃.
5. Rinse cells with DPBS (pH 7.4) and fix with formaldehyde-glutaraldehyde combined fixative for15 minat 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 scanning microscope.
Note:
1) Artificially introduced azide groups on the cell surface can be targeted by Cyanine5 DBCO for more than3 daysafter treatment with Ac4ManNAz.
2) Degree of Cyanine5 DBCO labeling is in a dose-dependent manner.
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体内研究
(In Vivo) | 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][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. Add Ac4ManNAz (50 μM, final concentration), incubate for3 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 dayspost 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.
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| 运输条件 | Room temperature in continental US; may vary elsewhere. |
| 储存方式 | -20°C, protect from light, stored under nitrogen *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen) |
| 溶解性数据 | In Vitro: DMSO : 125 mg/mL(134.55 mM;Need ultrasonic) 配制储备液 | 1 mM | 1.0764 mL | 5.3820 mL | 10.7639 mL | | 5 mM | 0.2153 mL | 1.0764 mL | 2.1528 mL | | 10 mM | 0.1076 mL | 0.5382 mL | 1.0764 mL |
*请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80℃, 6 months; -20℃, 1 month (protect from light, stored under nitrogen)。-80℃ 储存时，请在 6 个月内使用，-20℃ 储存时，请在 1 个月内使用。 |
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