Thermo Scientific NHS-Azide reagents are amine-reactive compounds that can be used to derivatize primary amines of proteins or amine-coated polymer surfaces for ligation to phosphine-modified molecules.
These NHS-ester compounds react to form covalent bonds with primary amines (e.g., side chain of lysine residues or aminosilane-coated surfaces). The azide (N3) group reacts with phosphine-labeled molecules by a mechanism known as Staudinger chemistry, enabling efficient and specific conjugation of derivatized molecules in biological samples. In combination with different phosphine-activated compounds, these NHS-Azide reagents facilitate a variety of labeling and crosslinking experimental strategies for investigating protein interactions and cellular pathways.
- Soluble – reagents easily dissolve in water-miscible solvents for subsequent dilution in aqueous reaction mixtures with cell lysates and other biological samples
- Compatible – reaction chemistry occurs effectively in simple buffer conditions; requires no accessory reagents such as copper or reducing agents
- Specific – NHS esters are specific for covalent attachment to primary amines (side chain of lysine or N-terminus of polypeptides)
- Chemoselective – azide and phosphine groups do not react or interfere with components of biological samples (bioorthogonal) but conjugate to one another with high efficiency
- Pegylated – choose the very short NHS-Azide or a variety whose spacer arm contains a 4-unit polyethylene glycol (PEG) group for increased solubility and reach
|Chemical structures of NHS-Azide Reagents. These reagents label proteins and molecules that contain primary amines (e.g., side-chain of lysine) via the NHS Ester Reaction Chemistry. Labeled molecules will contain an azide group (N3), which can be conjugated and detected using phosphine-activated probes via the Staudinger Ligation Reaction Chemistry.
Properties of NHS-Azide reagents. These compounds are identical except for the their overall length as a result of different numbers of polyethylene glycol groups (PEG units).
|Spacer arm length
||white to off-white powder
||colorless to pale yellow liquid
||-20°C with desiccant, protect from moisture and light, use only fresh solutions
||N-hydroxysuccinimide ester, reacts with primary amines at pH 7.0-9.0;
Azide, reacts with phosphine groups
Azide-Phosphine Reagent Guide
Review of Metabolic Labeling for Chemoselective Ligation
Review of Staudinger Ligation Reaction Chemistry
Superiority of Azide-Phosphine Chemistry
Azide-Sugars for Metabolic Labeling and Detection
NHS-Phosphine Staudinger Ligation Reagents
Phosphine-activated DyLight Fluors