Product Pathways - Cell Cycle / Checkpoint
Phospho-NPM (Thr199) Antibody #3541
|W IP IHC-P IF-IC||H M R||Endogenous||38||Rabbit|
Reactivity Key: H=Human M=Mouse R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Phospho-NPM (Thr199) Antibody detects endogenous levels of NPM only when phosphorylated at threonine 199.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Thr199 of human NPM. Antibodies are purified by protein A and peptide affinity chromatography.
Western blot analysis of extracts from HeLa cells synchronized at various stages of the cell cycle, using Phospho-NPM (Thr199) Antibody (upper) or NPM Antibody #3542 (lower).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using Phospho-NPM (Thr199) Antibody.
Confocal immunofluorescent analyis of HT-29 cells, untreated (left) or λ-phosphatase-treated (right), using Phospho-NPM (Thr199) Antibody (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Nucleophosmin (NPM; also known as B23, numatrin or NO38) is an abundant phosphoprotein primarily found in nucleoli. It has been implicated in several distinct cellular functions, including assembly and transport of ribosomes, cytoplasmic/nuclear trafficking, regulation of DNA polymerase α activity, centrosome duplication and molecular chaperoning activities (1,2). The NPM gene is also known for its fusion with the anaplastic lymphoma kinase (ALK) receptor tyrosine kinase. The NPM portion contributes to transformation by providing a dimerization domain, which results in activation of the fused kinase (3,4).
NPM associates with unduplicated centrosomes and is a direct substrate of Cdk2-cyclin E in centrosome duplication (4). Upon phosphorylation at Thr199 by Cdk2-cyclin E, NPM dissociates from centrosomes, and this dissociation is a prerequisite step for centrosome to initiate duplication (5).
- Okuda, M. et al. (2000) Cell 103, 127-140.
- Takemura, M. et al. (1999) J. Biochem. (Tokyo) 125, 904-909.
- Morris, S.W. et al. (1994) Science 263, 1281-1284.
- Bischof, D. et al. (1997) Mol. Cell. Biol. 17, 2312-2325.
- Tokuyama, Y. et al. (2001) J. Biol. Chem. 276, 21529-21537.
- Saavedra, H. I. et al. (2003) Inactivation of E2F3 results in centrosome amplification. Cancer Cell 3, 333-346. Applications: Western Blotting
- So, C.H. et al. (2012) J Biol Chem 287, 17088-99. Applications: Western Blotting
- Lin, C.Y. et al. (2010) Mol Biol Cell 21, 4409-17. Applications: Western Blotting
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For Research Use Only. Not For Use In Diagnostic Procedures.