Product Pathways - Cell Cycle / Checkpoint
Phospho-NPM (Thr199) Antibody #3541
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IP IHC-P IF-IC | H M R | Endogenous | 38 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species cross-reactivity is determined by Western blot.
Protocols
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 Blotting
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).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using Phospho-NPM (Thr199) Antibody.
IF-IC
Immunofluorescent analysis of NIH/3T3 cells, using Phospho-NPM (Thr199) Antibody (left) or the same antibody preincubated with phospho-NPM (Thr199) peptide (right).
Background
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.
Application References
- Saavedra, H. I. et al. (2003) Inactivation of E2F3 results in centrosome amplification. Cancer Cell 3, 333-346. Applications: Western Blotting
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Companion Products
- 3542 NPM Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
