About this item
The use of biotin for non-radioactive labeling of proteins and nucleic acids has now become an increasingly popular technique in life science research
G-Biosciences' HOOK™ Amine Reactive Biotin Reagents react with protein primary amines. Amines, lysine ε-amines and N-terminal α-amines, are the most abundant group in protein molecules and represent the most common target for biotinylation. For example, BSA contains 59 primary amines, of which up to 35 are available on the surface of the molecules and can be reacted with amine reactive esters.
The most widely used amine reactive biotinylation reagents are the water insoluble N-hydroxysuccinimide (NHS) esters or the water soluble N-hydroxysulfosuccinimide (sulfo-NHS) esters. The addition of a charged sulfonate (SO³⁻) on the N-hydroxysuccinimide ring of the sulfo-NHS esters results in their solubility in water (~10mM), but are not permeable to plasma membranes. The solubility and impermeability to plasma membranes makes them ideal for studying cell surface proteins as they will only react with the protein molecules on the outer surface of plasma membranes. The reaction of the NHS and sulfo-NHS esters with amines is virtually identical leading to the formation of an amide bond and release of NHS or sulfo-NHS.
G-Biosciences' HOOK™ Amine Reactive Biotin Reagents are available as the reagent alone or as part of a biotin labeling kit. The labeling kits are designed and supplied with all the necessary reagents, equipment and instructions necessary for optimal reaction conditions, efficient labeling, removal of unbound biotin, and quantification of biotin labeling, for the conjugation of biotin to protein primary amine groups. Each kit is supplied with 25 milligrams of the specific HOOK™ Amine Reactive Biotin Reagent, a specific Optimizer Buffer™ for enhanced conjugation, Spin-OUT™ columns for rapid (less than 10 minute) purification of labeled protein, and reagents to determine the amount of biotinylation. The kits are each designed for the coupling of 1 to 10 milligrams protein in 1 milliliter buffer, suitable for 10 couplings.
In addition, several of the more commonly used HOOK™ Biotin reagents are available in G-Biosciences' OneQuant™ format. The OneQuant™ format prevents loss of reagent due to repeated weighing, as each vial contains only 1 to 2 milligrams of the specific HOOK™ Biotin Reagent.
Several factors must be considered when coupling a biotin reagent to a protein to ensure a successful reaction. The primary consideration is the selection of the biotinylation reagent itself. G-Biosciences offers a wide range of biotin reagents with variations in their reactive groups, spacer arm lengths, solubility, membrane permeability and reversibility. All of these factors must be considered and are dependent on your protein/peptide.
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