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Anti phospho atr

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Anti-phospho-ATR is a primary antibody that recognizes ATR (Ataxia Telangiectasia and Rad3-related) protein when phosphorylated at specific serine or threonine residues. ATR is a key regulator of the DNA damage response pathway.

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Lab products found in correlation

Anti phospho atm, by Cell Signaling Technology (3 mentions) Anti phospho chk1, by Cell Signaling Technology (2 mentions) Anti atr, by Cell Signaling Technology (2 mentions) Anti chk1, by Cell Signaling Technology (2 mentions) Anti atm, by Cell Signaling Technology (2 mentions) Anti chk2, by Cell Signaling Technology (2 mentions) Anti sirt6, by Merck Group (1 mentions) Anti cdk2, by Cell Signaling Technology (1 mentions) Anti phospho chk2, by Cell Signaling Technology (1 mentions) Anti phospho akt, by Merck Group (1 mentions) Anti β tubulin, by Cell Signaling Technology (1 mentions) β actin, by Zhongshan Golden Bridge Biotechnology (1 mentions) Nitrocellulose membrane, by Cytiva (1 mentions) Anti cdc25b, by Cell Signaling Technology (1 mentions) Anti tubulin, by Merck Group (1 mentions) Anti p chk1, by Cell Signaling Technology (1 mentions) Anti ku70, by Cell Signaling Technology (1 mentions) Anti p chk2, by Cell Signaling Technology (1 mentions) Hrp conjugated antibody, by Abcam (1 mentions) Anti stat3, by Cell Signaling Technology (1 mentions)

6 protocols using anti phospho atr

1

Western Blot Antibody Profiling

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Western blot analysis was carried out as previously interpreted [23 (link), 24 (link)]. Primary antibodies in this experiment comprised of: anti-Sirt6 (Sigma), anti-phosphor-PI3 Kinase p110α, anti-phospho-AKT(Ser473), anti-total pan-AKT, anti phospho-mTOR, and anti-total pan-mTOR, anti-PTEN, anti-phospho-4EBP1, anti-FoxO1, anti-HIF-1α, anti-PARP [specific to the full-length (116 kDa) and the cleaved form (89 kDa) of PARP], anti-p27, anti-CDK2, anti-phospho-ATM, anti-phospho-ATR, anti-phospho-Chk1 (Ser345), anti-phospho-Chk2 (Thr68) and anti-β-tubulin (Cell Signaling Technology, MA, USA), β-actin (Zhongshan Goldenbridge, Beijing, China). Details are shown in Supplementary methods.
Yang J., Li Y., Zhang Y., Fang X., Chen N., Zhou X, & Wang X. (2020). Sirt6 promotes tumorigenesis and drug resistance of diffuse large B-cell lymphoma by mediating PI3K/Akt signaling. Journal of Experimental & Clinical Cancer Research : CR, 39, 142.
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2

Western Blot Analysis of DNA Damage Response

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For each sample, 1 × 107 cells were lysed as described previously.13 Equal amounts of proteins were separated on 7.5–13% SDS‐PAGE depending on the molecular sizes of the proteins, blotted onto a nitrocellulose membrane (Amersham Biosciences, Little Chalfon, UK) and blocked with 5% non‐fat drymilk in PBS/Tween (0.05% Tween‐20 in PBS). The following antibodies were used: anti‐ATM (D2E2), anti‐phospho‐ATM (10H11.E12), anti‐ATR, anti‐phospho‐ATR, anti‐Cdc25B, anti‐Cdc25C (5H9), anti‐CDK4, anti‐CDK6, anti‐phospho‐Chk1 (Ser317), anti‐phospho‐Chk1 (133D3, Ser345), anti‐Chk2, anti‐phospho‐Chk2 (Th68), anti‐cyclin D3 and anti‐cyclin E (HE12) from Cell Signaling Technology (Danvers, USA); anti‐phospho‐Cdc25A (Ser178) and anti‐phospho‐Cdc25A (Ser75) from Abgent (San Diego, CA, USA); anti‐Cdc25A (Clone DCS‐120 + DCS‐121) from NeoMarkers (Thermo Scientific, UK); anti‐Chk1 (FL‐475) from Santa Cruz Biotechnology (Heidelberg, Germany); anti‐γH2AX (JBW301, Ser139) from Millipore (Millerica, MA); anti‐γH2AX (Alexa‐Fluor‐488) from Biozol Diagnostics (Eching, Germany) and anti‐tubulin from Sigma (St. Louis, USA).
Neumann J., Yang Y., Köhler R., Giaisi M., Witzens‐Harig M., Liu D., Krammer P.H., Lin W, & Li‐Weber M. (2015). Mangrove dolabrane‐type of diterpenes tagalsins suppresses tumor growth via ROS‐mediated apoptosis and ATM/ATR–Chk1/Chk2‐regulated cell cycle arrest. International Journal of Cancer, 137(11), 2739-2748.
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3

Chidamide and MI-3 Effects on DNA Damage

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2×105/well cells were treated with chidamide in the absence or presence of and/or MI-3 for 48 h, and the subjected to western blot analysis using indicated primary antibodies and secondary HRP-conjugated antibodies (1:10,000, Abcam, Cambridge, UK). The primary antibodies included anti-caspase-3 (#9662S), anti-PARP (#9532S), anti-histone H3 (#4499S), anti-phospho-H3 (#53348S), anti-γH2A.X (#2577S), anti-RAD51 (#8875S), anti-KU70 (#4588S), anti-STAT3 (#9139 S), anti-Mcl-1 (#94296S), anti-phospho-p53 (#9286S), anti-p21(#2947S), anti-phospho-ATM (#5803S), anti-ATM (#2873S), anti-phospho-ATR (#2835S), anti-CHK1 (#2360), anti-CHK2 (#2662), anti-P-CHK1 (#2197S), and anti-P-CHK2 (#2348S) from Cell Signaling Technology (Boston, MA, USA). The primary antibodies were diluted with 5% fat-free milk-TBST. Anti-β-actin (1:1000, Cell Signaling Technology) was used as loading control. Blots were then detected using the ECL Western Blotting Detection Kit (GeneFlow, Staffordshire, UK).
Ye J., Zha J., Shi Y., Li Y., Yuan D., Chen Q., Lin F., Fang Z., Yu Y., Dai Y, & Xu B. (2019). Co-inhibition of HDAC and MLL-menin interaction targets MLL-rearranged acute myeloid leukemia cells via disruption of DNA damage checkpoint and DNA repair. Clinical Epigenetics, 11, 137.
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4

Monitoring DNA Damage Response Pathways

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The monoclonal antibodies used for this study included: anti-phospho-ATM (Ser 1981) (Cell Signaling Technology, Inc., Danvers, MA; Cat. #4526); anti-phospho-ATR (Cell Signaling Technology, Inc., Danvers, MA; Cat. #2853); anti-phospho-DNA PKcs (Ser2056) (Abcam, Cambridge, MA; Cat. ab18192); anti-phospho-Histone H2AX (Ser139) (Millipore, Mahopac, NY; Cat. #05-636); anti-8-OHdG (Abcam, Cambridge, MA; Cat. ab10802); LC3B (Invitrogen, Inc., Palo Alto, CA; Cat. L10382); NF-κB p65 (Cell Signaling Technology, Inc., Danvers, MA; Cat.4764S); iNOS (Cell Signaling Technology, Inc., Danvers, MA; Cat.13120).
Zhang B., Davidson M.M, & Hei T.K. (2014). Mitochondria regulate DNA damage and genomic instability induced by high LET radiation. Life sciences in space research, 1, 80-88.
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5

Western Blot Analysis of DNA Damage Signaling

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Total protein and phosphorylated protein levels were analyzed by western blot analysis. Briefly, following treatment, cells were lysed and whole-cell lysates resolved by SDS-PAGE (6 or 10%) and immunoblotted with the indicated antibodies. The following primary antibodies were used for immunoblotting: anti-AKT, anti-phospho-Akt (S473), anti-CHK1, anti-phospho-CHK1 (S345), anti-CHK2, anti-ATR, anti-phospho-ATR (S428), anti-ATM, anti-rabbit/mouse horseradish peroxidase-conjugated secondary antibodies (Cell Signaling Technology, Inc.), anti-Ku70, anti-phospho-CHK2 (T68) (Novus Biologicals LLC, Littleton, CO, USA), anti-phospho-ATM (S1981), anti-RAD51 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), and anti-β-actin (Millipore).
Dong J., Ren Y., Zhang T., Wang Z., Ling C.C., Li G.C., He F., Wang C, & Wen B. (2018). Inactivation of DNA-PK by knockdown DNA-PKcs or NU7441 impairs non-homologous end-joining of radiation-induced double strand break repair. Oncology Reports, 39(3), 912-920.
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6

Gastric Cancer Cell Line Characterization

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All human GC cell lines (BGC823, HGC27, MGC803, MKN45, and SGC7901), normal gastric cell line (GES-1) and human embryonic renal cell line 293FT were obtained from the American Type Culture Collection (ATCC, Beijing, China). All cell lines were tested mycoplasma-negative. MG132 and CHX were obtained from Sigma (Shanghai, China). Anti-ARIH2, anti-p21, anti-p27, anti-CDK1, anti-CDK2, anti-α-Tubulin, anti-HA, anti-SKP2, anti-RNF126 and anti-UHRF2 antibodies were purchased from Proteintech (Wuhan, China). Anti-MYC, anti-Flag, anti-phospho-ATR, anti-phospho-ATM, anti-ɣ-H2AX and anti-cleaved-caspase-3 antibodies were obtained from Cell Signaling Technology (Shanghai, China). Anti-Ki67 and anti-Bcl2 antibodies were purchased from BD Biosciences. All antibodies were diluted according to the manufacturer’s instructions.
Geng S., Peng W., Wang X., Hu X., Liang H., Hou J., Wang F., Zhao G., Lü M, & Cui H. (2022). ARIH2 regulates the proliferation, DNA damage and chemosensitivity of gastric cancer cells by reducing the stability of p21 via ubiquitination. Cell Death & Disease, 13(6), 564.
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