Immunological detection of phosphorylation.
Incorporation of phosphates into serine, threonine, and tyrosine acceptors in proteins is a common mechanism for regulating protein function. This unit presents protocols that use specific anti-phosphoamino acid (PAA) and anti-phosphoprotein antibodies to detect protein phosphorylation and protein kinase activity. Immunoblotting to detect protein phosphorylation using either anti-PAA or anti-phosphoprotein antibodies. This is a convenient method that usually yields impressive results. Phosphorylation can also be detected by immunoprecipitation followed by immunoblot analysis or by immunofluorescent staining; these methods are typically more complicated and time consuming. All three methods have been successfully used to detect protein phosphorylation with a wide variety of antibodies and most phosphorylated proteins.
Yao
Z
Z
The Weizmann Institute of Science, Rehovot, Israel.
Seger
R
R
D000818:Animals; D000911:Antibodies, Monoclonal; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D007158:Immunologic Techniques; D047468:Immunoprecipitation; D010766:Phosphorylation; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats
D000911:Antibodies, Monoclonal; D011506:Proteins