Background
Cy5.5-NHS ester is a near-infrared amine reactive dye for labeling amino groups of peptides, proteins and oligonucleotides. Cy5.5-NHS ester is suitable for fluorescence analysis when the background fluorescence interferes. It can also be used for in vivo imaging experiments. Cy5.5-NHS ester can be dissolved in DMF and DMSO, the maximum excitation light and maximum emission light are around 675 nm and 693 nm, respectively.[1]
In vitro and in vivo experiment indicated that Cy5.5-NHS ester is a nice near-infrared fluorescent dye, which could be used for tumor targeting as well as examining the effect of D-glucose or L-glucose on 2-NBDG accumulation. Moreover, Cy5.5-NHS ester reacts swiftly that the tumor/muscle contrast could be clearly visualized from 30 min up to 24 h pi of 0.5 nmol Cy5.5-NHS. The tumor uptake of Cy5.5-NHS reached a maximum 30 minutes pi. [2]
参考文献:
[1].Ju HR, et al. Non-invasive optical imaging of cathepsin B with activatable fluorogenic nanoprobes in various metastatic models. Biomaterials. 2014. 35: 2302-2311.
[2].Zhen C. et al. Near-Infrared Fluorescent Deoxyglucose Analog for Tumor Optical Imaging in Cell Culture and in Living Mice. Bioconjug Chem. 2006 ; 17(3): 662–669.
Cy5.5-NHS 酯是一种近红外胺活性染料，用于标记肽、蛋白质和寡核苷酸的氨基。 Cy5.5-NHS ester 适用于背景荧光干扰时的荧光分析。它还可用于体内成像实验。 Cy5.5-NHS酯可溶于DMF和DMSO，最大激发光和最大发射光分别在675 nm和693 nm左右。[1]
体外和体内实验表明，Cy5.5-NHS酯是一种很好的近红外荧光染料，可用于肿瘤靶向以及检测D-葡萄糖或L-葡萄糖对2- NBDG积累。此外，Cy5.5-NHS 酯反应迅速，在 0.5 nmol Cy5.5-NHS 的感染后 30 分钟至 24 小时内，肿瘤/肌肉对比清晰可见。 Cy5.5-NHS 的肿瘤摄取在感染后 30 分钟达到最大值。 [2]
Protocol
| Cell experiment [1]: |
|
Cell lines
|
U87MG
|
|
Preparation Method
|
U87MG cells (1 × 105) were cultured for 24 hours, then the cells were washed with PBS and then incubated at 37 °C in the presence of 100 nM Cy5.5-NHS ester for 10 minutes. After the incubation period, cells were washed three times with ice- cold PBS.
|
|
Reaction Conditions
|
The fluorescence signal of the cells was recorded using an Axiovert 200M fluorescence microscope equipped with a Cy5.5 filter set.
|
|
Applications
|
Cy5.5-NHS ester could be used for fluorescence microscopy imaging study with tumor cell lines such as U87MG, C6, A375M, B16F10 and MDAMB-435, as well as for examining the effect of D-glucose or L-glucose on 2-NBDG accumulation.
|
| Animal experiment [1]: |
|
Animal models
|
Female athymic nude mice (nu/nu), 4–6 weeks
|
|
Preparation Method
|
Mice were subcutaneously injected with 5 × 106 U87MG glioblastoma cells suspended in 100 μL of PBS. When the tumors reached 0.4–0.6 cm in diameter, the tumor bearing mice were then injected via tail vein with Cy5.5-NHS ester and subjected to optical imaging at various time points postinjection (pi).
|
|
Dosage form
|
0.5 nmol
|
|
Applications
|
Cy5.5-NHS ester could be took advantage of tumor targeting, and the tumor/muscle contrast was clearly visible as early as 30 min postinjection. The highest U87MG tumor/muscle ratio of 3.34 ± 0.23 were achieved 24 hours post-injection for Cy5.5-NHS.
|
|
参考文献:
[1]. Zhen C. et al. Near-Infrared Fluorescent Deoxyglucose Analog for Tumor Optical Imaging in Cell Culture and in Living Mice. Bioconjug Chem. 2006 ; 17(3): 662–669.
|
没有评价数据