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Aurora proteins are a family of serine/threonine kinases crucial for cell cycle control. Mutations of this kinase caused the formation of monopolar spindles surrounded by kinase, and the appearance of this was reminiscent of the Aurora borealis at the poles of the earth. Mammals express three closely related Aurora kinases named Aurora A, Aurora B, and Aurora C.
Image: HeLa cell cultures were stained with MO22159 antibody (green). The antibody stains spindle poles and mitotic spindles at anaphase and concentrates on the midbody between the two daughter cells during telophase. It is therefore a useful marker of dividing cells. Cells were counterstained with our chicken polyclonal antibody to Vimentin CH22108 red. Blue is a DNA stain.
Mammalian genomes encode 3 Aurora kinases named Aurora A, Aurora B, and Aurora C. All 3 contain a regulatory domain at the N terminus which is quite different between the molecules followed by a catalytic serine/threonine kinase domain which is almost identical between them. As a consequence antibodies raised against one Aurora family member frequently cross-react with other family members. Since there is a short C-terminal peptide which is also variable between the three molecules. Aurora A is first associated with centrosomes and then with spindle microtubules whereas Aurora B localizes to the spinal midzone and finally accumulates at the midbody. MO22159 is an excellent reagent for the visualization of mid bodies, centrosomes and spindles in dividing cells. The antibody was tested for binding to expressed human Aurora A, B and C and shown to react with both Aurora A and B, but not C (see Blot image)
Left: Western blot analysis of MO22159 in HeLa cells. Blot of HeLa cells treated with 100ng/ml nocodazole for 18 hours was probed with MO22159. Nocodazole is a microtubule depolymerizing agent which induces cells to halt at the G2/M phase and also induces Aurora A expression. The MO22159 monoclonal binds strongly to a band at about 46 kDa, which is Aurora A and also shows binding to a band at 38 kDa. Right: Blots of recombinant human Aurora A, B and C were probed with MCA-3H1, which binds to both Aurora A and B. We conclude that the 38 kDa band seen in the HeLa extract is Aurora B. This is also consisted with the immunocytochemical staining of both centrosomes and midbodies seen on HeLa cells.