Aurora A/B Substrate Antibody Sampler Kit #96882
Product Information
Kit Usage Information
Protocols
- 2187: Western Blotting, Immunoprecipitation (Magnetic), Immunofluorescence*
- 2914: Western Blotting, Immunofluorescence, Flow
- 3377: Western Blotting, Immunofluorescence, Flow
- 7074: Western Blotting
- 8842: Western Blotting, Immunofluorescence
- 9062: Western Blotting, Immunoprecipitation (Magnetic)
- 9713: Western Blotting, Immunofluorescence, Immunofluorescence, Flow
Product Description
Specificity / Sensitivity
Each antibody in the Aurora A/B Substrate Antibody Sampler Kit detects endogenous levels of its respective modification-specific target protein. Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb does not detect phosphorylated Ser10 when Lys9 is acetylated or methylated.
Source / Purification
Monoclonal and polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding their respective phospho-specific targets. Phospho-CENP-A (Ser7) Antibody is purified by peptide affinity chromatography and Phospho-Histone H3 (Ser28) Antibody is purified by protein A and peptide affinity chromatography.
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. Aurora kinase 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); research investigators have observed overexpression of these kinases 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, research studies report overexpression of Aurora C is detected in various cancer cell lines (6).
Transforming acid coiled-coil (TACC) proteins are a family of proteins characterized by a common coiled-coil motif of approximately 200 amino acids at the carboxy-terminal end (7). When phosphorylated at Ser558 by Aurora A, mammalian TACC3 is localized to mitotic spindles and increases microtubule stability (8,9).
Aurora A-dependent phosphorylation of CENP-A on Ser7 during prophase is required for proper targeting of Aurora B to the inner centromere in prometaphase, proper kinetochore/microtubule attachment, and proper alignment of chromosomes during mitosis (10). Aurora B also targets Ser7 on CENP-A, which in turn regulates Aurora B activity during cytokinesis (11). Aurora B phosphorylates both Ser10 and Ser28 on histone H3 in concordance with mitotic chromosome condensation (12).
Aurora A phosphorylation of Thr210 on PLK promotes mitotic entry following checkpoint-dependent cell cycle arrest (13). Autophosphorylation of conserved threonine residues Thr288 (Aurora A), Thr232 (Aurora B), and Thr195 (Aurora C) is required for Aurora kinase activity (14).
- Warner, S.L. et al. (2003) Mol Cancer Ther 2, 589-95.
- Katayama, H. et al. (2003) Cancer Metastasis Rev 22, 451-64.
- Andrews, P.D. et al. (2003) Curr Opin Cell Biol 15, 672-83.
- Pascreau, G. et al. (2003) Prog Cell Cycle Res 5, 369-74.
- Crosio, C. et al. (2002) Mol Cell Biol 22, 874-85.
- Kimura, M. et al. (1999) J Biol Chem 274, 7334-40.
- Gergely, F. et al. (2000) Proc Natl Acad Sci U S A 97, 14352-7.
- Kinoshita, K. et al. (2005) J Cell Biol 170, 1047-55.
- Schneider, L. et al. (2007) J Biol Chem 282, 29273-83.
- Kunitoku, N. et al. (2003) Dev Cell 5, 853-64.
- Zeitlin, S.G. et al. (2001) J Cell Biol 155, 1147-57.
- Goto, H. et al. (2002) Genes Cells 7, 11-7.
- Macůrek, L. et al. (2008) Nature 455, 119-23.
- Willems, E. et al. (2018) Cell Div 13, 7.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.