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Updated: Aug 2, 2026
Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions
Published on: August 2, 2015
Protocol for screening and validating antibodies specific to protein phosphorylation sites using a set of yeast
Monika Arbaciauskaite1, Azady Pirhanov2, Justin Paoloni1
1Department of Chemical and Biomolecular Engineering, University of Connecticut, Storrs, CT 06269, USA.
Abstract:
Developing antibodies with high specificity against post-translationally modified epitopes remains a challenge. Yeast biopanning is well suited in screening for high-specificity binders. Here, we present a protocol for screening and validating antibodies specific to protein phosphorylation sites using a set of yeast biopanning approaches. We describe steps for screening a yeast surface display library for antibodies and other binders. We then detail procedures for validating the antibodies found by analyzing their specificity through whole-well image analysis in 96-well plates. For complete details on the use and execution of this protocol, please refer to Arbaciauskaite et al.1.
