United Kingdom (UK)
Recombinant monoclonal antibody to Phosphovimentin. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 4A4.
UniProt Accession Number of Target Protein: P20152
Alternative Name(s) of Target: MPV55; HPV55; phosphorylated vimentin; vimentin phosphoSer55
Immunogen: The original antibody was generated by immunizing BDF1 mice intraperitoneally with a MPV55-KLH emulsified in complete Freund's adjuvant. MPV55-KLH is a mouse vimentin phosphopeptide MPV55 (Ser-Leu-Tyr-Ser-Ser-phosphoSef15-ProGly-Gly-Ala-Tyr-Cys) which was coupled to keyhole limpet hemocyanin (KLH) with m-maleimido-benzoyl-N-hydroxysuccinimide ester.
Specificity: This antibody specifically binds mouse and human vimentin phosphorylated at serine 55 by cdc2 kinase . This antibody is incapable of binding non-phosphorylated vimentin or with vimentin phosphorylated by other kinases.
Application Notes: This antibody was developed to investigate the role of intermediate filament (IF) protein phosphorylation by cdc2 kinase during mitosis. The binding specificity of 4A4 towards serine 55 phosphorylated vimentin was determined by ELISA. Western blotting indicated that this antibody reacted with vimentin phosphorylated by cdc2 kinase but not with non-phosphorylated vimentin or with vimentin phosphorylated by other kinases such as CAMP-dependent protein kinase, protein kinase C, or Ca2+-calmodulin-dependent protein kinase 11. Immunofluorescence and immunoelectron microscopy showed that vimentin Ser55 residues distributed in the entire cytoplasmic vimentin filament system are phosphorylated when the cells enter mitosis and dephosphorylated in cytokinesis (PMID: 7983050). This antibody was also used in the visualization of mitotic radial glial lineage cells in the developing rat brain which revealed different patterns of gliogenesis in the neocortex and cerebellum (PMID: 9633804). This antibody has been commonly used for the visualization and study of phosphorylation of vimentin during mitosis (PMID: 8601602, 8992382, 9565595).
Antibody first published in:
Tsujimura et al. Visualization and function of vimentin phosphorylation by cdc2 kinase during mitosis. J Biol Chem. (1994); 269(49):31097-106. PMID:7983050
Note on publication:
Decribes the generation of this antibody to determine the role of phosphorylation by cdc2 kinase during mitosis.