Biotinylation of immunoreactants

Percentage of amino groups substituted after reaction with biotin -hydroxysuccini-mide (BNHS) . From Guesdon et al. (1979) 

Molar ratio BNHS/amino group Sheep IgG GOase POase APase BGase 

Preparation of biotin derivatives active with groups other than amino groups (reactions 

Biotinyl-nitroanilide (BNA) is prepared (steps 1-6) by the mixed anhydride method (Section 12.3.1) its transformation to BDA (steps 7-10) should be performed immediately before use. 

Dissolve BNA in 2 ml DMF and add water until suspension becomes slightly turbid. 

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