Product Pathways - Cell Cycle / Checkpoint
Aurora Antibody Sampler Kit #3875
| Kit Includes | Quantity | Applications | Reactivity | MW (kDa) | Isotype |
|---|---|---|---|---|---|
| Phospho-Aurora A (Thr288) (C39D8) Rabbit mAb #3079 | 40 µl | W IF-IC | H | 48 | Rabbit IgG |
| Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11) XP® Rabbit mAb #2914 | 40 µl | W IF-IC F | H M R | 35, 40, 48 | Rabbit IgG |
| Aurora A/AIK (1G4) Rabbit mAb #4718 | 40 µl | W IP IF-IC | H Mk | 48 | Rabbit IgG |
| Aurora B/AIM1 Antibody #3094 | 40 µl | W IP F | H M R Mk | 40 | Rabbit |
| Anti-rabbit IgG, HRP-linked Antibody #7074 | 100 µl | Goat |
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 enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Each antibody in the Aurora Antibody Sampler Kit detects endogenous levels of its respective target protein and does not cross-react with other family members.
Western Blotting
Western blot analysis of extracts from HeLa, L929 and C6 cells, treated with 4 mM hydroxyurea for 20 hours or treated with 100 nM paclitaxel or 100 ng/ml nocodazole for 20 hours, using Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D11A13) XP® Rabbit mAb #2914 (upper) or Aurora B/AIM1 Antibody #3094 (lower).
Western Blotting
Western blot analysis of extracts from HeLa and HT29 cells, hydroxyurea or nocodazole treated, using Phospho-Aurora A (Thr288) (C39D8) Rabbit mAb #3079.
Western Blotting
Western blot analysis of extracts from Hela, NIH/3T3 and C6 cells, untreated or nocodazole-treated (50 ng/ml, 24 hrs), using Aurora B/AIM1 Antibody #3094.
Western Blotting
Western blot analysis of extracts from HeLa, 293 and COS cells, using Aurora A/AIK (1G4) Rabbit mAb #4718.
Description
The Aurora Antibody Sampler Kit provides an economical means to investigate the G2/M phase of the cell cycle. The kit contains enough primary and secondary antibodies to perform four Western blots with each antibody.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to the amino terminus of human Aurora B/AIM1. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of human Aurora A/AIK, residues around Thr288 of human Aurora A, or residues surrounding Thr232 of human Aurora B.
Background
Aurora kinases belong to a highly conserved family of mitotic serine/threonine kinases with three members identified among mammals: Aurora A, B, and 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 phase 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; Aurora B 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); overexpression of these 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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Protocols
- 2914:
- Flow, Immunofluorescence, Western Blotting
- 3079:
- Immunofluorescence, Western Blotting
- 3094:
- Flow, Immunoprecipitation, Western Blotting
- 4718:
- Immunofluorescence, Immunoprecipitation, Western Blotting
Companion Products
- 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
For Research Use Only. Not For Use In Diagnostic Procedures.
