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Custom Ordering Details: Product is assembled upon order to ensure maximum activity. United States: Please allow up to two weeks for your order to be processed and shipped. Outside of the United States: Please allow up to three weeks, depending on the country, for your order to be processed and shipped. When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

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REACTIVITY
Product Includes Volume Solution Color
Aurora A Ab Coated Microwells 96 tests
Aurora A Detection Ab 11 ml Green
Anti-mouse IgG, HRP-linked Antibody 11 ml Red
TMB Substrate 7004 11 ml Colorless
STOP Solution 7002 11 ml Colorless
ELISA Wash Buffer (20X) 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
Sealing Tape 2 sheets
Cell Lysis Buffer (10X) 9803 15 ml Yellowish

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.


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.


Entrez-Gene Id 6790
Swiss-Prot Acc. O14965


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