Antibody-based fluorescence detection of kinase activity on a peptide array.
Peptide-based microarrays allow for high-throughput identification of protein kinase substrates. However, current methods of detecting kinase activity require the use of radioisotopes. We have developed a novel fluorescence-based approach for quantitative detection of peptide phosphorylation on chip using fluorescently-labeled anti-phosphoserine and anti-phosphotyrosine antibodies. This method is sensitive, specific and extremely fast, presenting obvious advantages and may find wider uses in high-throughput kinase screenings.
Lesaicherre
Marie Laure
ML
Department of Chemistry, National University of Singapore, 3 Science Drive 3, 117543, Singapore, Singapore.
Uttamchandani
Mahesh
M
Chen
Grace Y J
GY
Yao
Shao Q
SQ
D000906:Antibodies / Q000276:immunology*; D005453:Fluorescence; D010766:Phosphorylation; D010768:Phosphoserine / Q000276:immunology / Q000378:metabolism; D010770:Phosphotransferases / Q000378:metabolism*; D019000:Phosphotyrosine / Q000276:immunology / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods*; D012680:Sensitivity and Specificity; D013379:Substrate Specificity; D013997:Time Factors
D000906:Antibodies; D010768:Phosphoserine; D019000:Phosphotyrosine; D010770:Phosphotransferases