Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Cell Cycle / Checkpoint

TPX2 (D2R5C) XP® Rabbit mAb #12245

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H Mk Endogenous 100 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  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

TPX2 (D2R5C) XP® Rabbit mAb recognizes endogenous levels of total TPX2 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys420 of human TPX2 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using TPX2 (D2R5C) XP® Rabbit mAb.

IP

IP

Immunoprecipitation of TPX2 from MCF7 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or TPX2 (D2R5C) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using TPX2 (D2R5C) XP® Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of MCF7 cells using TPX2 (D2R5C) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Background

The Ras family small GTPase Ran is involved in nuclear envelope formation, assembly of the mitotic spindle, and nuclear transport (1,2). TPX2, a target of active Ran (RanGTP), is a microtubule nucleating protein. TPX2 is inactive when bound to nuclear importin-alpha. RanGTP activity disrupts this interaction, relieving inhibition of TPX2 (3). TPX2 binding activates Aurora A kinase and promotes its localization to the mitotic spindle (4,5). DNA damage in mitosis leads to loss of interaction between Aurora A and TPX2 and inactivation of Aurora A kinase (6). TPX2 is highly expressed in pancreatic cancer cells, and knockdown of TPX2 expression in these cells is associated with increased sensitivity to paclitaxel (7).

  1. Quimby, B.B. and Dasso, M. (2003) Curr Opin Cell Biol 15, 338-44.
  2. Hetzer, M. et al. (2002) Nat Cell Biol 4, E177-84.
  3. Gruss, O.J. and Vernos, I. (2004) J Cell Biol 166, 949-55.
  4. Kufer, T.A. et al. (2002) J Cell Biol 158, 617-23.
  5. Bayliss, R. et al. (2004) Cell Cycle 3, 404-7.
  6. Bhatia, P. et al. (2010) Cell Cycle 9, 4592-9.
  7. Warner, S.L. et al. (2009) Clin Cancer Res 15, 6519-28.

Application References

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Companion Products

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

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