Product Pathways - Cell Cycle / Checkpoint
Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11) Rabbit mAb #2914
| Applications | Reactivity | Sensitivity | MW (kDa) | Source | Isotype |
|---|---|---|---|---|---|
| W IF-IC F | H M R | Endogenous | 35,40,48 | Rabbit | IgG |
Applications Key:
W=Western Blotting
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11) Rabbit mAb detects endogenous levels of Aurora A/B/C only when phosphorylated at either Thr288, Thr232 or Thr198 respectively.
Source / Purification
Monoclonal antibody is produced by immunizing rabbits with a synthetic peptide (KLH-coupled) derived from residues surrounding Thr232 of human Aurora B.
Flow Cytometry
Flow cytometric analysis of untreated Jurkat cells using Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11) Rabbit mAb compared to propidium iodide (DNA content). The boxed population indicates phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198)-positive cells.
IF-IC
Confocal immunofluorescent analysis of HT-1080 cells using Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11) Rabbit mAb (green), β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red), and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (blue).
Western blot analysis of extracts from HeLa, L929 and C6 cells, treated with 4 mM hydroxyurea for 20 hours to induce G1/S phase or treated with 100 nM paclitaxel or 100 ng/ml nocodazole for 20 hours to induce G2/M phase, using Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) D11A13 Rabbit mAb (upper) or Aurora B/AIM1 Antibody #3094 (lower).
Background
Aurora kinases belong to a highly conserved family of mitotic serine/threonine kinases with three members identified among mammals: Aurora A, Aurora B and Aurora C (1,2). Studies on the temporal expression pattern and subcellular localization of Aurora kinases in mitotic cells suggest an association with mitotic structure. Their functional influences span from G2 to cytokinesis and may be involved in key cell cycle events such as centrosome duplication, chromosome bi-orientation and segregation, cleavage furrow positioning and ingression (3). Aurora A is detected at the centrosomes, along mitotic spindle microtubules and in the cytoplasm of mitotically proliferating cells. Aurora A protein levels are low during G1 and S phases and peak during the G2/M phase of the cell cycle. Phosphorylation of Aurora A at Thr288 in its catalytic domain increases kinase activity. Aurora A is involved in centrosome separation, maturation and spindle assembly and stability. Expression of Aurora B protein also peaks during the G2/M phase of the cell cycle, while kinase activity peaks at the transition from metaphase to the end of mitosis. Aurora B associates with chromosomes during prophase prior to relocalizing to the spindle at anaphase. Aurora B regulates chromosome segregation through the control of microtubule-kinetochore attachment and cytokinesis. Expression of both Aurora A and Aurora B during the G2/M phase transition is tightly coordinated with histone H3 phosphorylation (4,5), while overexpression of both kinases is seen in a variety of human cancers (2,4). Aurora C localizes to the centrosome from anaphase to cytokinesis and both mRNA and protein levels peak during G2/M phase. Although typical Aurora C expression is limited to the testis, overexpression of Aurora C is detected in various cancer cell lines (6).
- Warner, S.L. et al. (2003) Mol. Cancer Ther. 2, 589-595.
- Katayama , H. et al. (2003) Cancer Metastasis Rev. 22, 451-464.
- Andrews, P.D. et al. (2003) Curr. Opin. Cell Biol. 15, 672-683.
- Pascreau, G. et al. (2003) Prog. Cell Cycle Res. 5, 369-374.
- Crosio, C. et al. (2002) Mol. Cell. Biol. 22, 874-885.
- Kimura, M. et al. (1999) J. Biol. Chem. 274, 7334-7340.
Application References
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Companion Products
- 3079 Phospho-Aurora A (Thr288) (C39D8) Rabbit mAb
- 4718 Aurora A/AIK (1G4) Rabbit mAb
- 3092 Aurora A/AIK Antibody
- 3094 Aurora B/AIM1 Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked 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
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
