Biotinylation of proteins

The following protocol is a suggested method for the biotinylation of proteins with either NHS-LC-biotin or sulfo-NHS-LC-biotin. 

Biotinylation of proteins on blots using the diazonium derivative of p-aminobenzoyl biocytin. 

Biotinylation of Proteins on Blots Using the Diazonium Derivative of p-Aminobenzoyl Biocytin... 

Cull, M. G. and Schatz, P. J. (2000) Biotinylation of proteins in vivo and in vitro using small peptide tags. Methods Enzymol. 326, 430-440. 

Chapman-Smith, A. and Cronan, J. E. (1999) The enzymatic biotinylation of proteins a posttranslational modification of exceptional specificity. Trends Biochem. Sci. 24, 359-363. 

Lue, R. Y, Zhu, Q., Li, D. (2004). Site-specific peptide immobilization strategies for the rapid detection of kinase activity on microarrays. Site-specific immobilization of biotinylated proteins for protein microarray analysis. Enzymatic profiling system in a small-molecule microarray. Intein-mediated biotinylation of proteins and its application in a protein microarray. Developing site-specific immobilization strategies of peptides in a microarray. Methods Mol. Biol. 278, 191-204. 

Reagents for biotinylation of proteins and peptides are commercially available and allow the fast and efficient derivatization of antibodies and enzymes without the loss of enzyme or antibody activity. Several reagents are also available for the biotinylation of sulfhydryl groups aldehydes, nucleic acids and carbohydrates. Hence, in addition to proteins and peptides, a variety of antigens can be labeled with biotin. 

Selective biotinylation of proteins (e.g., phosphoproteins, etc.) Dependent on the specificity of labeling excellent purification of targeted proteins... 

Biotinylation of protein. Mix antigen solution in PBS (pH 7-8.5) Avith sulfo-NHS-biotin solution in proportions of 25 p,g antigen xg sulfo-NHS-biotin. Incubate the mixture at room temperature for 30 min. 

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