Blocking and detection chemistries affect antibody performance on reverse phase protein arrays
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Updated: Jul 4, 2026

Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples
Published on: May 4, 2012
Kristi L H Ambroz1, Yonghong Zhang, Amy Schutz-Geschwender
1LI-COR Biosciences, 4647 Superior Street Lincoln, NE 68504, USA. kristi.ambroz@licor.com
Abstract:
Antibody specificity is critical for RP protein arrays (RPA). The effects of blocking and detection chemistries on antibody specificity were evaluated for Western blots and RPA. Blocking buffers significantly affected nonspecific banding on Western blots, with corresponding effects on arrays. Tyramide signal amplification (TSA) increased both specific and nonspecific signals on Westerns and arrays, masking the expected gradations in signal intensity. These results suggest that consistent blocking and detection conditions should be used for antibody validation and subsequent RPA experiments.
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