Anti aurora a

Manufactured by Abcam

Sourced in United Kingdom

Anti-Aurora A is a primary antibody that binds to Aurora A, a serine/threonine protein kinase involved in cell cycle regulation. This product is intended for research use only.

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4 protocols using anti aurora a

Protein Extraction and Western Blot Analysis

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Cells were harvested in TLB lysis buffer supplemented with the Complete C protease and phosphatase inhibitors mix (Roche, Mannheim, Germany). Quantification was made with Bradford ready to use reagent (Biorad, Hercules, CA). Total cell protein (10 μg) was separated by 12 % SDS- PAGE, transferred to nitrocellulose membrane (Biorad, Hercules, CA) and incubated with primary antibodies overnight at 4°C followed by incubation with goat-anti-mouse (Invitrogen, Carlsbad, CA, USA) or goat-anti-rabbit (Invitrogen, Carlsbad, CA, USA) HRP tagged secondary antibodies. Bands were visualized with Lumigen on Amersham Hyperfilm (GE Healthcare, Fairfield, CT, USA). Primary antibodies used are listed below: anti-WEE1 (NP_001137448.1) (Abcam, Cambridge, UK), anti-WIP1 (NP_003611.1) (Abcam, Cambridge, UK), anti-pCHK1 Ser345 (Cell Signaling, Boston MA, USA), anti- γH2AX (Genetex, Irvine, CA), anti-p21 (NP_000380.1) (Genetex, Irvine, CA), anti-MYT1 (NP_004526.1) (Genetex, Irvine, CA), anti-Aurora A (NP_003591.2) (Abcam, Cambridge, UK), anti-CDC25B (NP_001274445.1) (Genetex, Irvine, CA) and anti-PLK1 (NP_005021.2) (Abcam, Cambridge, UK); anti-GAPDH (NP_001243728.1) was used as loading control (Genetex, Irvine, CA).

Rodríguez A., Torres L., Juárez U., Sosa D., Azpeitia E., Teresa B.G., Cortés E., Ortíz R., Salazar A.M., Ostrosky-Wegman P., Mendoza L, & Frías S. (2015). Fanconi anemia cells with unrepaired DNA damage activate components of the checkpoint recovery process. Theoretical Biology & Medical Modelling, 12, 19.

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Esophageal Cancer Tissue Expression

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Esophageal cancer tissue array(cat#: ES481, ES482, ES807)were purcased from Yingchao Biotech, Xi'an, China. Specimens were incubated with anti-Aurora-A (Abcam; 1∶700) and anti-PAK7 (MBL; 1∶50) at 4°C overnight following the antigen retrieval in pH9.0 EDTA buffer in an autoclave for 2 min. All immunostaining experiments were assessed by an experienced pathologist. The protein expression within the cancer tissue was evaluated and categorized according to the percentagee and intensity of positive cells (tissues with no or weak staining in ≤5% of cells as “−/+”; with moderate staining in 6% to 25% of cells as “+”; with strong staining in 26% to 50% of cells as “++”; and with strong staining in ≥50% of cells as “+++”). Tumors were then further grouped into weak (−/+ and +) and strong (++ and +++) expression of each protein.

He S., Feng M., Liu M., Yang S., Yan S., Zhang W., Wang Z., Hu C., Xu Q., Chen L., Zhu H, & Xu N. (2014). P21-Activated Kinase 7 Mediates Cisplatin-Resistance of Esophageal Squamous Carcinoma Cells with Aurora-A Overexpression. PLoS ONE, 9(12), e113989.

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Western Blot Analysis of Protein Markers

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Total cell lysates were prepared with 1% SDS and 10 mM Tris buffer (pH 7.4), and separated by SDS-PAGE. After transfer to a nitrocellulose membrane, the proteins were immunoblotted with different antibodies overnight at 4°C. The following antibodies were used: anti-ubiquitin (catalog # 3933) (1:1000), anti-p21 (catalog #: 2947) (1:500), anti-survivin (catalog #: 2808) (1:2000), anti-cleaved PARP (catalog #: 9546) (1:300), and anti-cyclin B1 (catalog #: 12231) (1:1000), all from Cell Signaling Technology (Boston, MA); anti-aurora A from Abcam (Cambridge, MA) (catalog #: ab190367) ; and anti-β-actin (catalog #: sc-81178) (1:3000) from Santa Cruz Biotechnology (Dallas, TX). Anti-human β-actin was used as a loading control. The membranes were incubated with the appropriate HRP-conjugated IgG (anti-rabbit antibody at 1:3000 dilution, Cell Signaling Technology, Danvers, MA, USA, or anti-mouse antibody at 1:10 000 dilution, Santa Cruz Biotechnology). An ECL assay (Thermo Scientific, Rockford, IL, USA) was used to detect the proteins.

Zhang L., Boufraqech M., Lake R, & Kebebew E. (2016). Carfilzomib potentiates CUDC-101-induced apoptosis in anaplastic thyroid cancer. Oncotarget, 7(13), 16517-16528.

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Dissecting STAU1 Domains via Plasmids

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Plasmids coding for STAU1 Δ2 -HA3, STAU1 Δ3 -HA3, STAU1 Δ4 -HA3, STAU1 Δ5 -HA3, STAU1 ΔTBD -HA3, STAU1 3*-4* -HA3, STAU1 Δ25 -HA3, STAU1 Δ37 -HA3, as well as RBD2-HA3 and RBD4-TBD-HA3 were previously described (Luo et al., 2002; Martel et al., 2010; Chatel-Chaix et al., 2008; Wickham et al., 1999; Boulay et al., 2014) . Monoclonal (1/1000) and rabbit (1/1000) anti-STAU1 antibodies were previously described (Dugre-Brisson et al., 2005; Rao et al., 2019) , respectively. Anti-β-actin (A5441, clone AC-74. 1/5000), anti-STAU2 (HPA0191551/500), anti-α-tubulin (T6074, batch number 023M4813. 1/40000), and anti-HA (H6908, batch number 115M4872v. 1/1000) antibodies were purchased from Sigma-Aldrich. Anti-aurora A (30925, batch number 2. 1/2000), anti-α-tubulin (for IF, ab18251, batch number GR201260-1. 1/40000) and anti-histone H3 (ab1791, batch number GR204148-1. 1/3000) antibodies were purchased from Abcam. Anti-calnexin (sc-11397, batch number C1214. 1/1000) antibody was obtained from Santa Cruz. Anti-RPS6 (2212, batch number 4. 1/1000) was purchased from Cell Signaling. Anti-RPL26, (GTX101833. 1/1000) and anti-STAU1 (for IF, GTX106566. 1/200) was obtained from Genetex. Goat polyclonal anti-mouse (p0447, bach number 20051789. 1/3000) and antirabbit (p0448, batch number 20017525. 1/5000) antibodies were purchased from Dako.

Hassine S., Bonnet‐Magnaval F., Benoit-Bouvrette L., Doran B., Ghram M., Bouthillette M., Lécuyer É., & DesGroseillers L. (2020). Staufen1 localizes to the mitotic spindle and controls the transport of RNA populations to the spindle.

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