Application

Western Blotting Analysis: A representative lot detected both the endogenous Bora and a FLAG-tagged Bora exogenously expressed in HEK293T cells. Downregulation of both the endogenous and FLAG-tagged Bora was seen following replication stress induction by camptothecin (Cat. No. 208925) or UV treatment of HEK293T cells (Qin, B., et al. (2013). J. Biol. Chem. 288(22):16139-16144).
Western Blotting Analysis: A representative lot detected FLAG-tagged wild-type and T501A Bora exogenously expressed in U2OS transfectants. Only the wild-type, but not the T501A mutant, was seen downregulated upon UV irradiation of the transfected U2OS cells (Qin, B., et al. (2013). J. Biol. Chem. 288(22):16139-16144).
Immunoprecipitation Analysis: A representative lot immunorecipitated significantly higher amount of ubiquitinated Bora from UV-irradiated HEK293T cells pre-treated with the proteasome inhibitor MG-132 than UV-treated cells without MG-132 (Qin, B., et al. (2013). J. Biol. Chem. 288(22):16139-16144).
Immunoprecipitation Analysis: A representative lot immunorecipitated Bora with enhanced Thr501 phosphorylation from UV irradiated HEK293T cells (Qin, B., et al. (2013). J. Biol. Chem. 288(22):16139-16144).

Anti-Bora Antibody is an antibody against Bora for use in Western Blotting, Immunoprecipitation.

General description

Protein aurora borealis (UniProt Q6PGQ7; also known as HsBora) is encoded by the BORA (also known as C13orf34) gene (Gene ID 79866) in human. Originally identified as a co-factor for Aurora A-dependent asymmetrical cell divisions, Bora is an adaptor/scaffold protein that plays an important role in mitosis by mediating Aurora A-dependent Plk1 activation, and Bora depletion abolishes both Plk1 and Cdk1 activation. Cellular Bora activity is negarively regulated by Plk1- and Cdk1-mediated phosphorylation. Cdk1 initially phosphorylates Bora on S252, which primes Bora for Plk1-catalyzed S497 and T501 phosphorylation, leading to SCFβTrCP-mediated Bora ubiquitination and subsequent proteasomal degradation. The negative feed-back regulation of Bora by the kinase it activates is important, as uncontrolled Bora overexpression perturbs Aurora A and Plk1 localization, leading to lengthened metaphase. Similar to the counter regulation between Bora and Cdk1/Plk1 in somatic cells, Bora degradation is reported to be regulated via phosphorylation by Cdk1 and Plx1 in xenopus ooctyes. Cdk1 is reported to phosphorylate Bora on Thr52, which blocks its degradation. Fertilization induced calcium surge jump-starts the oscillations of the embryonic cycles by inducing Calcineurin-catalyzed Bora pThr52 dephosphorylation, leading to Bora degradation.

Immunogen

Epitope: N-terminal.

GST-tagged recombinant human Bora N-terminal fragment.

Other Notes

Concentration: Please refer to lot specific datasheet.

Quality

Evaluated by Western Blotting in HEK293 cell lysate.

Western Blotting Analysis: An 1:500 dilution of this antibody detected Bora in 10 µg of HEK293 cell lysate.

Specificity

This rabbit polyclonal antiserum reacts with unmodified as well as postranslationally modified (ubiquitination and phosphorylation) Bora (Qin, B., et al. (2013). J. Biol. Chem. 288(22):16139-16144). Expected to react with both spliced isoforms of human Bora reported by UniProt (Q6PGQ7).

Target description

~58 kDa observed. 61.20 kDa (isoform 1) and 53.44 kDa (isoform 2) calculated.