Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

INCENP (P240) Antibody #2807

Applications Reactivity Sensitivity MW (kDa) Source
W IF-IC F H Endogenous 140 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
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

INCENP (P240) Antibody detects endogenous levels of total INCENP protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids surrounding Pro240 of human INCENP. Antibodies are purified by peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts of Jurkat and MOLT4 cells using INCENP (P240) Antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, using INCENP (P240) Antibody (blue) compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of mitotic HeLa cells during metaphase (left) or anaphase (right) using INCENP (P240) Antibody (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).


Background

INCENP (inner centromere protein antigens 135 kDa, 155 kDa) is a chromosomal passenger protein crucial for multiple events that mediate chromosome separation during mitosis (1). At prophase INCENP is associated with chromatin whereas during prometaphase and metaphase it translocates to the inner centromere (1). Depletion of INCENP results in aberrant chromosome alignment at the metaphase plate, incomplete chromosome separation, and disruption of proper spindle formation and cytokinesis (2). INCENP is part of the chromosomal passenger complex that also contains Aurora B, borealin and survivin (2). Aurora B and INCENP are mutually dependent on each other for proper localization (3), and in Drosophila cells and C.elegans embryos that lack INCENP or survivin, Aurora B cannot organize the kinetochores and the midbody (4,5). Phosphorylation on INCENP by CDK1 on Thr59 and Thr388 leads to the association of INCENP with Plk1, another important regulator of mitotic entry and exit (6). Interaction of INCENP with Plk1 is necessary for recruitment of Plk1 to the kinetochores, and the metaphase to anaphase transition (6). Interactions have also been reported between INCENP and heterochromatin protein 1α (HP1) (7) and β-tubulin (8).

  1. Carmena, M. and Earnshaw, W.C. (2006) Nat Cell Biol 8, 110-1.
  2. Carmena, M. and Earnshaw, W.C. (2003) Nat Rev Mol Cell Biol 4, 842-54.
  3. Kaitna, S. et al. (2000) Curr Biol 10, 1172-81.
  4. Speliotes, E.K. et al. (2000) Mol Cell 6, 211-23.
  5. Adams, R.R. et al. (2001) J Cell Biol 153, 865-80.
  6. Goto, H. et al. (2006) Nat Cell Biol 8, 180-7.
  7. Ainsztein, A.M. et al. (1998) J Cell Biol 143, 1763-74.
  8. Wheatley, S.P. et al. (2001) Exp Cell Res 262, 122-7.

Application References

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Companion Products

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

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