Thermo Scientific NHS-Phosphine is an amine-reactive compound that can be used to derivatize primary amines of proteins or amine-coated polymer surfaces for ligation to azide-tagged molecules.
This NHS-ester compound reacts to form covalent bonds with primary amines (e.g., side chain of lysine residues or aminosilane-coated surfaces). The phosphine group (PPh2) reacts with azide-labeled molecules by a mechanism known as Staudinger chemistry, enabling efficient and specific conjugation of derivatized molecules in biological samples. In combination with different azide-tagged compounds, this NHS-Phosphine reagent facilitates a variety of labeling and crosslinking experimental strategies for investigating protein interactions and cellular pathways.
- Soluble – reagent easily dissolves 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
|Chemical structure of NHS-activated (amine-reactive) phosphine ligation reagent. This reagent labels proteins and molecules that contain primary amines (e.g., side-chain of lysine) via NHS Ester Reaction Chemistry. Labeled molecules will contain a phosphine group (P3), which can be conjugated to azide-activated molecules via the Staudinger Ligation Reaction Chemistry.
Properties of NHS-Phosphine.
|Spacer arm length
||Yellow to yellow-orange powder
||-20°C with desiccant, protect from moisture, use only fresh solutions
||N-hydroxysuccinimide ester, reacts with primary amines at pH 7.0-9.0;
Phosphine, reacts with azide 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-Azide Staudinger Ligation Reagents
Phosphine-activated DyLight Fluors