A novel approach to enhance antibody sensitivity and specificity by peptide cross-linking

作者:Namiki Takeshi; Valencia Julio C; Hall Matthew D; Hearing Vincent J*
来源:Analytical Biochemistry, 2008, 383(2): 265-269.
DOI:10.1016/j.ab.2008.08.024

摘要

Most current techniques employed to improve antigen-antibody signals in Western blotting and in immunohistochemistry rely on sample processing prior to staining (e.g., microwaving) or using a more robust reporter (e.g., a secondary antibody with biotin-streptavidin). We have developed and optimized a new approach intended to stabilize the complexes formed between antigens and their respective primary antibodies by cupric ions at high pH. This technique improves the affinity and lowers cross-reactivity with nonspecific bands of approximately 20% of antibodies tested (5/25). Here we report that this method can enhance antigen-antibody specificity and can improve the utility of some poorly reactive primary antibodies. Published by Elsevier Inc.

  • 出版日期2008-12-15