Analysis of phosphorylation of unlabeled proteins.
Phosphorylation of unlabeled proteins can be detected using immunological or enzymatic techniques. Anti-phosphotyrosine antibodies are used with immunoblots to detect tyrosine phosphorylation. This unit provides a protocol for detection of antibodies by 125I-labeled protein A or enhanced chemiluminescence (ECL). Detection of enzymatic dephosphorylation using a general or phosphoamino acid-specific phosphatase and subsequent SDS-PAGE mobility shift (or a change in activity) is also described.
Sefton
B M
BM
The Salk Institute, San Diego, California, USA.
D000906:Antibodies / Q000302:isolation & purification; D015151:Immunoblotting; D007457:Iodine Radioisotopes; D049449:Luminescence; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010766:Phosphorylation; D011506:Proteins / Q000276:immunology / Q000378:metabolism; D014443:Tyrosine / Q000276:immunology / Q000378:metabolism*
D000906:Antibodies; D007457:Iodine Radioisotopes; D011506:Proteins; D014443:Tyrosine; D010744:Phosphoric Monoester Hydrolases