Cell Lysis and Western Blot Analysis

Cell lysis and WB were performed as described in reference 62 (link). Antibodies were as follows: rabbit anti-TPX2 (1:1,000; Novus Biologicals), mouse anti-Aurora-A (0.5 µg/ml; BD Transduction Laboratories), rabbit anti-pAurora-A (Thr288) (3079 clone C39D8, 1:100; Cell Signaling Technology, Inc.), and mouse anti-GAPDH (SC-32233, 1:1,000; Santa Cruz Biotechnology). HRP-conjugated secondary antibodies (Bio-Rad Laboratories S.r.l.) were revealed using the Clarity Western ECL Substrate (Bio-Rad Laboratories S.r.l.).

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Asteriti I.A., Polverino F., Stagni V., Sterbini V., Ascanelli C., Naso F.D., Mastrangelo A., Rosa A., Paiardini A., Lindon C, & Guarguaglini G. (2023). AurkA nuclear localization is promoted by TPX2 and counteracted by protein degradation. Life Science Alliance, 6(5), e202201726.

Publication 2023

rabbit anti-TPX2 mouse anti-Aurora-A mouse anti-GAPDH (SC-32233, HRP-conjugated secondary antibodies Clarity Western ECL Substrate

Antibodies Aurora a Biologicals Cell Clone Gapdh Mouse Novus Rabbit Tpx2

Corresponding Organization : National Research Council

Other organizations : Fondazione Santa Lucia, University of Cambridge, Italian Institute of Technology

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein levels of TPX2, Aurora-A, and phosphorylated Aurora-A (Thr288)

control variables

GAPDH protein levels (used as a loading control)

controls

Positive control: None mentioned
Negative control: None mentioned

Annotations

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