Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total Aurora A Sandwich ELISA Antibody Pair #7117

Kit Includes Volume Cap Color
Aurora A Capture Ab (100X) 0.4 ml Pink
Aurora A Detection Ab(100X) 0.4 ml Blue
Anti-mouse IgG, HRP-linked Antibody 0.04 ml Yellow

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Important Ordering Details

Product is assembled upon order to ensure maximum activity. Domestic: Please allow up to two weeks for your order to be processed and shipped. International: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.

Protocols

Description

CST's PathScan® Total Aurora A Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Total Aurora A Sandwich ELISA Kit #7116. Capture and Detection Antibodies (100X stocks) and HRP-conjugated Secondary Antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Aurora A Rabbit Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by Aurora A Mouse Detection Antibody and HRP-conjugated Secondary Antibody. HRP substrate (TMB) is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of Aurora A protein.*Antibodies in this kit are custom formulations specific to the kit.

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Sandwich ELISA

Sandwich ELISA

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

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).

  1. Warner, S.L. et al. (2003) Mol. Cancer Ther. 2, 589-595.
  2. Katayama , H. et al. (2003) Cancer Metastasis Rev. 22, 451-464.
  3. Andrews, P.D. et al. (2003) Curr. Opin. Cell Biol. 15, 672-683.
  4. Pascreau, G. et al. (2003) Prog. Cell Cycle Res. 5, 369-374.
  5. Crosio, C. et al. (2002) Mol. Cell. Biol. 22, 874-885.
  6. Kimura, M. et al. (1999) J. Biol. Chem. 274, 7334-7340.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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