Rabbit anti phospho atr ser428

Manufactured by Cell Signaling Technology

Sourced in United States

Rabbit anti-phospho-ATR (Ser428) is a primary antibody that recognizes the phosphorylated form of the Ataxia Telangiectasia and Rad3-related (ATR) protein at serine 428. ATR is a key regulator of the DNA damage response pathway.

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2 protocols using rabbit anti phospho atr ser428

Western Blotting and Cell Fractionation

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Western blotting was performed as already described^{55 (link)}. The following antibodies were used: mouse anti-GLI1 (#2643), mouse anti-cyclin A2 (#4656), rabbit anti-BCL2 (#2876), rabbit anti-BAX (#2772), rabbit anti-cyclin B1 (#12231), rabbit anti-PARP-1 (#9532), rabbit anti-phospho-ATR (Ser428) (#2853), rabbit anti-phospho-CHK1 (Ser345) (#2348), rabbit anti-phospho-CDC2 (Tyr15) (#4539), rabbit anti-phospho-H2A.X (Ser139) (#9718), rabbit anti-phospho-Histone H3 (Ser10) (#3377), rabbit anti-phospho-WEE1 (Ser642) (#4910) (Cell Signaling Technology, Danvers, MA, USA), rabbit anti-CDC2 (sc-954), mouse anti-Myc (sc-40), mouse anti-HSP90 (sc-13119) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti-SMO (ST1718) (Merck Millipore, Burlington, MA, USA). Chemiluminescent detection was used. Cell fractionation was performed as previously described^{56 (link)}. The following antibodies were used: mouse anti-GLI1 (#2643) (Cell Signaling Technology), goat anti-fibrillarin (D-14), and goat anti-GAPDH (V-18) (Santa Cruz Biotechnology).

Pietrobono S., Santini R., Gagliardi S., Dapporto F., Colecchia D., Chiariello M., Leone C., Valoti M., Manetti F., Petricci E., Taddei M, & Stecca B. (2018). Targeted inhibition of Hedgehog-GLI signaling by novel acylguanidine derivatives inhibits melanoma cell growth by inducing replication stress and mitotic catastrophe. Cell Death & Disease, 9(2), 142.

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Immunoblotting Analysis of DNA Damage Response

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Protein extraction and separation on SDS-PAGE gels and immunoblotting was performed as previously described (5). The isolation and irradiation of separate populations from CA1 and Luc4 cell lines was repeated twice. Each set of samples (two sets) was immunoblotted twice. Primary antibodies used were rabbit anti-phospho-CHK2 (Thr68) (Cell Signaling), rabbit anti-phospho-ATM (Ser1981) (Cell Signaling), rabbit anti-phospho-ATR (Ser428) (Cell Signaling), rabbit anti-RAD50 (Cell Signaling), rabbit anti-RAD52 (Cell Signaling), rabbit anti-phospho-BRCA1 (Ser1524), rabbit anti-XLF (Cell Signaling), and mouse anti-β-Actin (Sigma). Secondary antibodies used were polyclonal rabbit anti-mouse immunoglobulin/HRP (DakoCytomation), and polyclonal goat anti-rabbit immunoglobulin/HRP (DakoCytomation). Densitometry was performed on scanned immunoblot images using the ImageJ gel analysis tool [21 ]. The gel analysis tool was used to obtain the absolute intensity for each experimental protein band and its corresponding protein loading (β-Actin) control band.

Gemenetzidis E., Gammon L., Biddle A., Emich H, & Mackenzie I.C. (2015). Invasive oral cancer stem cells display resistance to ionising radiation. Oncotarget, 6(41), 43964-43977.

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