Immunoprecipitation of Aurora-A and PFKFB3

Immunoprecipitation was performed as previously described [22 (link)]. Briefly, cells were lysed with IP lysis buffer (Thermo Scientific, USA) containing the cocktail. Cell lysates were incubated with anti-Aurora-A antibody or PFKFB3 overnight at 4 °C, followed by immunoprecipitation with protein A + G agarose beads (Thermo Scientific, USA) for 2 h at 4 °C with shaking and washed 3 times with IP lysis buffer. The immunoprecipitate was eluted and analyzed by western blotting.

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Jingtai Z., Linfei H., Yuyang Q., Ning K., Xinwei Y., Xin W., Xianhui R., Dongmei H., Weiwei Y., Xiangrui M., Tianze Z., Wei W, & Xiangqian Z. (2023). Targeting Aurora-A inhibits tumor progression and sensitizes thyroid carcinoma to Sorafenib by decreasing PFKFB3-mediated glycolysis. Cell Death & Disease, 14(3), 224.

Publication 2023

IP lysis buffer protein A + G agarose beads

Agarose Anti antibody Aurora a Buffer Cell Immunoprecipitation Protein a Protein g

Corresponding Organization :

Other organizations : Tianjin Medical University Cancer Institute and Hospital, Nankai University, Tianjin Medical University, Tianjin First Center Hospital

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Variable analysis

independent variables

Anti-Aurora-A antibody
PFKFB3

dependent variables

Immunoprecipitation
Western blotting

control variables

IP lysis buffer (Thermo Scientific, USA) containing the cocktail
Protein A + G agarose beads (Thermo Scientific, USA)

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