Cat. No.
MABL-2694
Application
ELISA, WB, IF
Isotype
Engineer antibody
Species Reactivity
Human, Mouse
Clone No.
4A4
From
Recombinant Antibody
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.
Alternative Names
MPV55; HPV55; phosphorylated vimentin; vimentin phosphoSer55
UniProt
P20152
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.
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
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.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
