Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-NPM (Thr95) Antibody #3517

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-IC F H (M) (R) (Mk) Endogenous 38 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
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 (Thr95) Antibody detects endogenous levels of NPM only when phosphorylated at Thr95.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Thr95 of human NPM. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts of HeLa cells, untreated or nocodazole-treated, using Phospho-NPM (Thr95) Antibody (upper) or NPM Antibody #3542 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells using Phospho-NPM (Thr95) Antibody versus propidium iodide (DNA content). The box indicates phospho-NPM positive cells.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells untreated (left) and λ phosphatase-treated (right), using NPM (Thr95) phosphate Antibody (green). Actin filaments have been labeled using DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (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).

  1. Okuda, M. et al. (2000) Cell 103, 127-140.
  2. Takemura, M. et al. (1999) J. Biochem. (Tokyo) 125, 904-909.
  3. Morris, S.W. et al. (1994) Science 263, 1281-1284.
  4. Bischof, D. et al. (1997) Mol. Cell. Biol. 17, 2312-2325.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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