英文名:Cyanine5-Azide,Cy5-N3, Cy5 Azide
中文名:菁染料CY5叠氮, 菁染料CY5叠氮
CAS:1782950-80-4
性 状:深蓝色粉末/溶液
分子量:601.22
分子式:C35H45ClN6O
溶解度:溶于有机溶剂(DMSO、DMF、二氯甲烷),水溶性低。
纯度:>95%
新研博美产品描述:点击化学用5-氰基叠氮化物标记试剂,在DMSO中以10 mM溶液形式提供,以固体形式提供。该叠氮化物可溶于有机溶剂(例如DMSO、DMF),因此应使用少量有机共溶剂进行标记反应。该叠氮化物可用于在水与有机溶剂的混合物中标记炔烃修饰的生物分子。二甲基亚砜溶液可用于生物结合。也可提供该试剂的水溶性磺化版本。氰基5是Cy5的类似物,与各种仪器兼容的最常用荧光团之一。
结构式:
We recommend using the following general protocol for Click chemistry labeling of alkyne-modified oligonucleotides with azides produced by Lumiprobe Corp. The auxiliary reagents can be ordered at Lumiprobe Corp.
- Calculate the volumes of reagents required for Click chemistry labeling using the table below. Prepare the required stock solutions (see Appendix).
Reagent Final concentration in the mixture Stock solution concentration Oligonucleotide, alkyne-modified Varies (20 — 200 uM) varies Azide 1.5 x (oligonucleotide concentration) 10 mM in DMSO DMSO 50 vol % — Ascorbic acid 0.5 mM 5 mM in water Cu-TBTA complex 0.5 mM 10 mM in 55 vol % DMSO - Dissolve alkyne-modified oligonucleotide or DNA in water in a pressure-tight vial.
- Add 2M triethylammonium acetate buffer, pH 7.0, to final concentration 0.2 M.
- Add DMSO, and vortex.
- Add azide stock solution (10 mM in DMSO), and vortex.
- Add the required volume of 5mM Ascorbic Acid Stock solution to the mixture, and vortex briefly.
- Degass the solution by bubbling inert gas in it for 30 seconds. Nitrogen, argon, or helium can be used.
- Add the required amount of 10 mM Copper (II)-TBTA Stock in 55% DMSO to the mixture. Flush the vial with inert gas and close the cap.
- Vortex the mixture thoroughly. If significant precipitation of azide is observed, heat the vial for 3 minutes at 80°C, and vortex.
- Keep at room temperature overnight.
- Precipitate the conjugate with acetone (for oligonucleotides) or with ethanol (for DNA). Add at least 4-fold volume of acetone to the mixture (If the volume of the mixture is large, split in several vials). Mix thoroughly and keep at −20°C for 20 minutes.
- Centrifuge at 10000 rpm for 10 minutes.
- Discard the supernatant.
- Wash the pellet with acetone (1 mL), centrifuge at 10000 rpm for 10 minutes.
- Discard the supernatant, dry the pellet, and purify the conjugate by RP-HPLC or PAGE.