Anti-Aurora A + B Kinase Antibody [5A12] (A85399)

Western blot analysis of different cell lysates and recombinant protein solutions using Anti-Aurora A + B Kinase Antibody [5A12] (A85399), in green. Left: Cells were treated with 100 ng/ml of nicodazol for 6 hours. The lanes contain samples of: [1] Protein standards, [2] HeLa cells, [3] A72 cells, [4] NBL6 cells, and [5] KR158 cells. Bands at 46kDa and 38kDa correspond to Aurora A and Aurora B proteins respectively. Right: Western blot of full-length recombinant human Aurora A, B and C, probed with Anti-Aurora A + B Kinase Antibody [5A12] (A85399). The antibody binds to Aurora A and Aurora B, but not to the closely related Aurora C, shown by the bands in lanes A and B.

HeLa cell cultures were stained with Anti-Aurora A/B Kinase Antibody (green). The antibody stains spindle poles and mitotic spindles at anaphase (top 2 panels) and concentrates on the midbody between the two daughter cells during telophase (bottom 2 panels). It is therefore a useful marker of dividing cells. Cells were counterstained with Anti-Vimentin Antibody (A85421 | red). Blue is a DNA stain.

Left: Western blot analysis of Anti-Aurora A/B Kinase Antibody in HeLa cells. Blot of HeLa cells treated with 100ng/ml nocodazole for 18 hours was probed with Anti-Aurora A/B Kinase Antibody. Nocodazole is a microtubule polymerization inhibitor which induces cells to halt at the G2/M phase and also induces Aurora A expression. The Anti-Aurora A/B Kinase Antibody 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 Anti-Aurora A/B Kinase Antibody. We conclude that this antibody reacts with the Aurora A and Aurora B kinases and that the 38 kDa band seen in the HeLa extract is Aurora B. This is also consistent with staining of both centrosomes and midbodies seen on HeLa cells.