NHS-Chromogenic-PEGj-Biotin

Proteins biotinylated with this reagent will have a characteristic absorbance band at 354 nm, which can be used to determine accurately the number of biotin groups per molecule. No other biotinylation compound has such built-in quantification capability. This feature eliminates the need to consume conjugate by doing a HABA assay to test for the level of biotin incorporation (Chapter 23, Section 7). 

The following protocol is adapted from the manufacturers recommendations. 

Dissolve a protein or other amine-containing molecule to be biotinylated in 0.1M sodium phosphate, 0.15M NaCl, pH 7.2—7.5, at a concentration of l-10mg/ml. Note 

NHS ester Chromogenic Hydrophilic PEG3 Biotin group 

Chromogenic biotinylated molecule via amide bond linkage 

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