ABSTRACT
Phosphate addition is a posttranslational modification of proteins, and this modification can affect the activity and other properties of intracellular proteins. Different animal species can be used to generate phosphosite-specific antibodies as either polyclonals or monoclonals, and each approach offers its own benefits and disadvantages. The validation of phosphosite-specific antibodies requires multiple techniques and tactics to demonstrate their specificity. These antibodies can be used in arrays, flow cytometry, and imaging platforms. The specificity of phosphosite-specific antibodies is vital for their use in proteomics and profiling of disease.
Subject(s)
Amino Acids/immunology , Antibodies/immunology , Antibody Specificity/immunology , Phosphoproteins/immunology , Amino Acids/metabolism , Animals , Antibody Formation , Blotting, Western , Cell Surface Display Techniques , Drug Discovery/methods , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Gene Expression Profiling , Humans , Hybridomas/immunology , Hybridomas/metabolism , Immunohistochemistry , Mice , Phosphoproteins/genetics , Phosphoproteins/metabolism , Phosphorylation , Protein Array Analysis/methods , Reproducibility of ResultsABSTRACT
Protein phosphorylation is a universal key posttranslational modification that affects the activity and other properties of intracellular proteins. Phosphosite-specific antibodies can be produced as polyclonals or monoclonals in different animal species, and each approach offers its own benefits and disadvantages. The validation of phosphosite-specific antibodies requires multiple techniques and tactics to demonstrate their specificity. These antibodies can be used in arrays, flow cytometry, and imaging platforms. The specificity of phosphosite-specific antibodies is key for their use in proteomics and profiling of disease.
