Product Pathways - Cell Cycle / Checkpoint
Phospho-NPM (Thr199) Antibody #3541
PhosphoSitePlus® protein, site, and accession data: NPM1
| 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. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
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
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).
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
- 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
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
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
For Research Use Only. Not For Use In Diagnostic Procedures.
