期刊
PROTEOMICS
卷 8, 期 12, 页码 2379-2383出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/pmic.200700676
关键词
antibody; fluorescence detection; protein arrays; reverse phase protein array; Western blotting
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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