An Overview of Phosphor Specific Antibody

1. An Overview of Phosphor Specific Antibody " _— ff? F= VIAN ™\ \ What is a phosphorylated antibody? Antibodies phosphorylation phosphorylation and dephosphorylation of the substrate and the change antibody binding type are used to phosphorylated, thereby studying changes immunogen is phosphorylated, the produced against dephosphorylation a phosphorylation site in the so state that of and of a substrate, the in investigate whether the substrate protein antibody prepared is in activity. After the with it is called phosphorylated antibody. The significance of phosphorylated antibody Post-translational modifications are essential for protein biological functions and play an important role in many phenomena. Among them, protein phosphorylation common important post-translational modifications. cellular activities in the body, such as signal transduction, cell cycle, proliferation and metabolism, growth and development, and cancer mechanisms. Phosphorylation addition of phosphate (PO4) groups to proteins under the action of kinases, most commonly on serine (S), threonine (T) or tyrosine (Y) residues. Since phosphorylated proteins can play a complex quantification of phosphorylated proteins at the cellular level is essential for deciphering and understanding cellular functions. key regulatory mechanisms of life is one of the most It participates in almost all it is closely related to many biological problems is usually the role, accurate and precise

2. The difference between phosphor specific antibodies antibodies and ordinary 1. The detection of phosphorylated antibody active state (phosphorylation). is based on the protein in an 2. Phospho-antibodies are only designed for phosphorylation sites. specific polyclonal antibody, which has some characteristics of monoclonal antibodies. It is a site- 3. expression protein-> purification-> immunize animal-> harvest antibody. Synthesis of common antibody (polyclonal antibody): prokaryotic 4. Synthesis of Phosphorylated antibody (phosphor specific antibodies): artificial synthesis phosphorylation antibodies. of peptides containing phosphorylation haptens-> sites-> animals-> artificial harvest in vitro -> link immunize 5. A large part of phosphorylated proteins are transcription-related factors. Therefore, phosphorylated antibodies and common antibodies are used to detect the same protein, in situ detection may have different cell localizations. Phosphorylated transcription factor moves nucleus. from the cytoplasm into the 6. directly content of the protein and cannot know antibodies measure the level of phosphorylation of the protein, focusing on understanding an active state of the protein. In general, the experimental results of phosphorylated proteins are more related to molecular functions. Ordinary antibodies measure the its active state. Phosphorylated 7. Phosphorylated antibodies are mainly prepared for phosphorylation sites, and detect the protein's changes when the cells are stimulated (whether the phosphorylation level is increased or decreased). Ordinary antibodies (mainly prepared for peptides at non-phosphorylated sites) mainly detect the expression of the protein in tissues, cells, tissue distribution, or localization in cells, phosphorylation levels. and can be used as an internal reference for changes in The application of phosphorylated antibodies 1. The level of protein phosphorylation many signaling pathway proteins responses by changing their phosphorylation level after certain stimulation, but their protein content may not change much. is often related to protein activity: are caused by a series of cellular

3. 2. Phosphorylated antibody: Prepared for specific phosphorylation sites, and detect changes in the protein's functional state (enhanced or weakened) when the cell is stimulated. 3. Non-phosphorylated antibody: It mainly detects the expression of the protein, tissue distribution or cell localization in tissues or cells, and can be used as an internal reference for changes in phosphorylation level.