Description
N-
Hydroxysuccinimide (NHS) activates carboxylic acid groups (on biotin) to facilitate coupling reactions. Biotin-
NHS is a compound used to attach biotin to primary amines under alkaline conditions (pH~8-
9). For example, lysines on the surface of proteins, including antibodies, are ideal targets for biotinylation with this compound.
Chemical Properties
White Solid
Uses
Biotin-NHS is used in the biotinylation of proteins and peptides. Couples with primary amine in the pH range 6.5-8.5.
Uses
amine reactive biotinylation reagent
Uses
Amino reactive biotin reagent for preparing biotinylated surfaces or polypeptides.
reaction suitability
reaction type: Biotinylations
Synthesis
GENERAL STEPS: To a mixed solution of D-biotin (0.82 mmol, 200 mg) and N-hydroxybutanediimide (0.89 mmol, 102 mg) was added 3-(ethyliminomethylamino)-N,N-dimethylpropan-1-amine hydrochloride (0.96 mmol, 184 mg). The reaction mixture was stirred at room temperature overnight and then concentrated to give a white solid. The crude product was mixed with isopropanol, heated to 70 °C and cooled. The pure product was obtained by filtration as a white powder in 91% yield (0.75 mmol, 255 mg).
Purification Methods
Recrystallise the ester from refluxing isoPrOH and dry it in a vacuum over P2O5 + KOH. [Jasiewicz et al. Exp Cell Biol 100 213 1976.]
References
[1] Journal of the American Chemical Society, 2007, vol. 129, # 45, p. 13987 - 13996
[2] New Journal of Chemistry, 2013, vol. 37, # 11, p. 3762 - 3769
[3] Organic Letters, 2013, vol. 15, # 16, p. 4130 - 4133
[4] Journal of Materials Chemistry, 2004, vol. 14, # 17, p. 2638 - 2642
[5] Beilstein Journal of Organic Chemistry, 2015, vol. 11, p. 784 - 791
