BiotinAlk can be used to biotinylate azide containing metabolic labels using bioorthogonal chemistry.12 The biotin handle can then be used in combination with various (strept)avidin constructs to enable the isolation, staining and fluorescent labeling of the biotinylated constructs.
Mode of action
BiotinAlk contains a biotin affinity handle, a hydrophilic polyethylene glycol (PEG) linker and an alkyne group. The alkyne group can be reacted selectively with azide containing biomolecules in complex biological samples on either live of fixed cells. This reaction is catalyzed by a copper catalyst which is cytotoxic at high concentrations and long reaction times. Hence, we recommend the use of copper binding ligands which accelerate the azide alkyne cycloaddition reaction, allows for the use of a lower copper concentration and short reaction times thereby reducing cytotoxicity. Excess BiotinAlk is washed away after the labeling and a (strept)avidin reagent of choice can be used to enable the isolation of visualization of biotinylated constructs.
Biotinylation of azide containing biomolecules on live or fixed cells. The biotin handle can then be used with a (strept)avidin reagent of choice to enable the isolation or visualization of biotinylated contructs.
BiotinAlk is soluble in PBS and is added to cell culture from a stock solution.