Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-PBK/TOPK (Thr9) Antibody #4941

Applications Reactivity Sensitivity MW (kDa) Source
W H Endogenous 40 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human
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-PBK/TOPK (Thr9) Antibody detects endogenous levels of PBK/TOPK only when phosphorylated at threonine 9.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to amino acids around Thr9 of human PBK/TOPK. Antibodies are purified by protein A and affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HT29 cells, untreated or nocodazole-treated (50ng/ml), using Phospho-PBK/TOPK (Thr9) Antibody (upper) or PBK/TOPK Antibody #4942 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated Jurkat cells, using Phospho-PBK/TOPK (Thr9) Antibody versus propidium iodide (DNA content). The boxed population indicates phospho-PBK/TOPK (Thr9)-positive cells.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HT-29 cells, untreated (left) or λ phosphatase-treated (right), using Phospho-PBK/TOPK (Thr9) Antibody (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Background

PBK/TOPK is a serine/threonine kinase that is phosphorylated and active during mitosis (1). PBK/TOPK is composed of kinase subdomains and a carboxy-terminal PDZ-Binding domain, which is thought to interact with the tumor suppressor protein hDlg (1). Increased PBK/TOPK expression has been observed in highly proliferative malignant cell lines, and PBK/TOPK expression is strongly downregulated during terminal differentiation of HL-60 leukemic cells (2,3). PMA-induced kinase activity toward PBK/TOPK has been observed (4), and cdc2/cyclinB has been shown to phosphorylate PBK/TOPK in vitro, presumably at Thr9 (1). Potential substrates of PBK/TOPK include p38 MAPK and c-Myc (3,4).

  1. Gaudet, S. et al. (2000) Proc. Natl. Acad. Sci. U S A 97, 5167-5172.
  2. Simons-Evelyn, M. et al. (2001) Blood Cells Mol. Dis. 27, 825-829.
  3. Nandi, A. et al. (2004) Blood Cells Mol. Dis. 32, 240-245.
  4. Abe, Y. et al. (2000) J. Biol. Chem. 276, 21525-21531.
  5. Matsumoto, S. et al. (2004) Biochem Biophys Res Commun. 325, 997-1004.

Application References

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Companion Products

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

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