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7116
PathScan® Total Aurora A Sandwich ELISA Kit
ELISA Kits
ELISA Kit

PathScan® Total Aurora A Sandwich ELISA Kit #7116

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PathScan® Total Aurora A Sandwich ELISA Kit: Image 1

Figure 1. Induced phosphorylation of Aurora A in paclitaxel-treated Hela cells lysed in the presence of phosphatase inhibitors (phospho lysate) is detected by PathScan® Phospho-Aurora A (Thr288) Sandwich ELISA Kit #7114 (upper, right). In contrast, a low level of phospho-Aurora A protein is detected in paclitaxel-treated HeLa cells lysed without addition of phosphatase inhibitors to the lysis buffer (nonphospho lysate). Similar levels of Aurora A protein from either nonphospho or phospho lysates are detected by PathScan® Total Aurora A Sandwich ELISA Kit #7116 (upper, left). The absorbance readings at 450 nm are shown in the top figure, while the corresponding Western blots using Aurora A Antibody #3092 (left panel) or Phospho-Aurora A (Thr288) Rabbit mAb #3079 (right panel) are shown in the bottom figure.

PathScan® Total Aurora A Sandwich ELISA Kit: Image 2

Figure 2: The relationship between protein concentration of phospho or nonphospho Aurora A lysates and the absorbance at 450 nm is shown. HeLa cells (85% confluence) treated with paclitaxel (100 nM) for 20 hours were harvested and then lysed in the absence or presence of phosphatase inhibitor.

To Purchase # 7116

Important Ordering Details

Custom Ordering Details: Product is assembled to order. Please allow up to three business days for your order to be processed.

Supporting Data

REACTIVITY

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Description

The PathScan® Total Aurora A Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total Aurora A protein. An Aurora A Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, Aurora A (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, an Aurora A Mouse Detection Antibody is added to detect the captured phospho and nonphospho Aurora A protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of total Aurora A protein.

Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

CST's PathScan® Total Aurora A Sandwich ELISA Kit #7116 detects endogenous levels of total Aurora A proteins. As shown in Figure 1, a significant induction of Phospho-Aurora A (Thr288) can be detected in HeLa cells treated with paclitaxel using the Phospho-Aurora A (Thr288) Sandwich ELISA Kit #7114. These high levels are abolished when paclitaxel-treated HeLa cells were lysed without addition of phosphatase inhibitors* to the lysis buffer. The levels of total Aurora A protein (either phospho or nonphospho) detected by PathScan® Total Aurora A Sandwich ELISA Kit #7116 remain unchanged during the G2/M phase of the cell cycle. Western analysis of protein captured in microwells coated with Aurora A antibody shows a single band corresponding to Aurora A protein (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

* Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate and Na3VO4.

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).

  1. Warner, S.L. et al. (2003) Mol Cancer Ther 2, 589-95.
  2. Katayama, H. et al. (2003) Cancer Metastasis Rev 22, 451-64.
  3. Andrews, P.D. et al. (2003) Curr Opin Cell Biol 15, 672-83.
  4. Pascreau, G. et al. (2003) Prog Cell Cycle Res 5, 369-74.
  5. Crosio, C. et al. (2002) Mol Cell Biol 22, 874-85.
  6. Kimura, M. et al. (1999) J Biol Chem 274, 7334-40.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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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's 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's 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.

For Research Use Only. Not For Use In Diagnostic Procedures.
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PathScan is a trademark of Cell Signaling Technology, Inc.