SH2 domain-based tyrosine phosphorylation array.
Tyrosine phosphorylation plays a major role in intracellular signal transduction pathways. Phosphorylated tyrosine residues initiate signaling pathways by recruiting proteins containing Src homology-2 (SH2) domains. Herein is described a high-throughput assay to detect interactions between phosphorylated proteins and human SH2 domains, in order to profile the genome-wide phosphorylation status of cells under a variety of biological conditions. The SH2 domain array was prepared by immobilizing 115 SH2 domain proteins on a membrane. The array assay is straightforward with no expensive equipment or radioactivity required: the cell lysate is incubated with the array membrane, and the signal is measured using a chemiluminescence-based detection system.
Jiang
Xin
X
Panomics, Inc., Herts, 6519 Dumbarton Cr., Freemont, CA, USA.
Roth
Lesile
L
Han
Stephanie
S
Li
Xianqiang
X
D000818:Animals; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D007202:Indicators and Reagents; D008322:Mammals; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics*; D040081:Protein Array Analysis; D011505:Protein-Tyrosine Kinases / Q000737:chemistry* / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry; D018909:src Homology Domains / Q000235:genetics*
D017931:DNA Primers; D018076:DNA, Complementary; D007202:Indicators and Reagents; D010750:Phosphoproteins; D011994:Recombinant Proteins; D005982:Glutathione Transferase; D011505:Protein-Tyrosine Kinases