Production of a site- and phosphorylation state-specific antibody.
Protein phosphorylation plays important roles in various aspects of cellular events. Visualization of site-specific phosphorylation in cells is of great importance not only to analyze spatial and temporal distribution but also to investigate biological function. Now, site- and phosphorylation state-specific antibodies are widely utilized as the most powerful tools for these analyses. This protocol details a method to produce the polyclonal version of such an antibody by immunizing a synthetic phosphopeptide corresponding to a protein phosphorylated at targeted site(s). This protocol is also applicable to the production of other types of antibodies, which specifically recognize the site-specific modification, such as acetylation, methylation and proteolysis. The protocol can be completed in 2-3 months.
Goto
Hidemasa
H
Division of Biochemistry, Aichi Cancer Center Research Institute, Nagoya, Aichi 464-8681, Japan.
Inagaki
Masaki
M
D045163:Antibodies, Phospho-Specific / Q000737:chemistry* / Q000302:isolation & purification; D007150:Immunohistochemistry; D010748:Phosphopeptides / Q000276:immunology; D010766:Phosphorylation; D011506:Proteins / Q000032:analysis / Q000378:metabolism
D045163:Antibodies, Phospho-Specific; D010748:Phosphopeptides; D011506:Proteins