Protein expression analysis by Western blot

RIPA buffer augmented with protease and phosphatase inhibitors was administered for cell lysis (14 (link)). The proteins were separated by SDS-PAGE and then transferred to nitrocellulose blotting membranes (GE Healthcare Life Science, Germany). The spots were examined with antibodies against PARP (Cell Signaling Technology, CST, Danvers, MA, USA, 9542), vinculin (Abcam, Cambridge, MA, USA, ab129002), GAPDH (HUABIO, Woburn, MA, USA, M1310-2), ATM (Santa Cruz Biotechnology, Dallas, TX, USA, sc135663), p-ATM (Ser1981) (CST, 5883), H2AX (Santa Cruz Biotechnology, sc517336), p-H2AX (Ser139) (Santa Cruz Biotechnology, sc517348), CHK1 (Santa Cruz Biotechnology, sc8408), p-CHK1 (Ser345) (CST, 2348), P53 (Santa Cruz Biotechnology, sc126), p-P53 (Ser15) (CST, 9286), ATR (Santa Cruz Biotechnology, sc515173), p-ATR (Ser428) (CST, 2853), FANCD2 (Santa Cruz Biotechnology, sc20022), and RAD51 (Santa Cruz Biotechnology, sc398587).

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Wang K.N., Hu Y., Han L.L., Zhao S.S., Song C., Sun S.W., Lv H.Y., Jiang N.N., Xv L.Z., Zhao Z.W, & Li M. (2022). Salvia chinensis Benth Inhibits Triple-Negative Breast Cancer Progression by Inducing the DNA Damage Pathway. Frontiers in Oncology, 12, 882784.

Publication 2022

nitrocellulose blotting membranes vinculin GAPDH p-H2AX FANCD2 RAD51

Antibodies Buffer Cell Fancd2 Gapdh Inhibitors Membranes Nitrocellulose Phosphatase Protease Proteins Ripa Sds page Spots Vinculin

Corresponding Organization :

Other organizations : Dalian Medical University, Second Affiliated Hospital of Dalian Medical University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression and phosphorylation levels of PARP, vinculin, GAPDH, ATM, p-ATM (Ser1981), H2AX, p-H2AX (Ser139), CHK1, p-CHK1 (Ser345), P53, p-P53 (Ser15), ATR, p-ATR (Ser428), FANCD2, and RAD51

control variables

None explicitly mentioned

controls

Positive controls: Not specified
Negative controls: Not specified

Annotations

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