5-TAMRA Picolyl Azide
5-TAMRA Picolyl Azide is an advanced fluorescent probe that incorporates a copper-chelating motif to raise the effective concentration of Cu(I) at the reaction site. The chelating effect of picolyl azide moiety to raise the effective concentration of copper at the reaction site is so great that it leads to an increase of signal intensity of up to 40-fold. In addition, the use of 5-TAMRA Picolyl Azide instead of conventional 5-TAMRA Azide allows for at least a tenfold reduction in the concentration of the copper catalyst without sacrificing the efficiency of labeling.
Although the mixed isomers of 5(6)-TAMRA is a preferred, routinely used red fluorescent dye for staining proteins, labeling peptides and nucleotides with TAMRA mixed isomers might be troublesome due to significant signal broadening in HPLC purification. Peptides and nucleotides labeled with a single isomer TAMRA usually give better resolution in HPLC purification that is often required in the conjugation processes.
In summary, the introduction of a picolyl moiety into an azide probe leads to a substantial increase in the sensitivity of alkyne detection. This will be of special value for the detection of low abundance targets or where significant increase in signal intensity is desired.
|Flow cytometry laser line||532 or 555 nm|
|Microscopy laser line||532 or 555 nm|
|Spectrally similar dyes||Alexa Fluor® 546, Atto™ 543, CF™ 555 Dye, DyLight™549|
|Solubility||DMSO, DMF, MeOH|
|Shipping conditions||Ambient temperature|
1. Jiang, H., et al. (2014). Monitoring Dynamic Glycosylation in Vivo Using Supersensitive Click Chemistry. Bioconjugate Chem., 25(4): 698-706.
2. Uttamapinant, C., et al. (2012). Fast, Cell-Compatible Click Chemistry with Copper-Chelating Azides for Biomolecular Labeling. Angew. Chem. Int. Ed,. 51(24): 5852–5856.
3. Gaebler, A., et al. (2016). A highly sensitive protocol for microscopy of alkyne lipids and fluorescently tagged or immunostained proteins. J. Lipid. Res.. DOI: 10.1194/jlr.D070565.
|MB 543 Picolyl Azide||A bright, water-soluble and pH-insensitive orange fluorescence-emitting probe.|
|Click Chemistry Reaction Buffer Kit||All-inclusive, click reaction enabling kit for protein labeling.|
|THPTA||A water-soluble ligand for copper-catalyzed azide-alkyne cycloadditions (CuAAC).|