Phospho atms1981

Manufactured by Abcam

Phospho-ATMs1981 is a lab equipment product that serves as a tool for the detection and quantification of phosphorylation of the ATM serine/threonine kinase at the 1981 serine residue. It can be used in various research applications involving the study of cell signaling pathways and DNA damage response mechanisms.

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Lab products found in correlation

Phospho p53 s15, by Cell Signaling Technology (2 mentions) γh2ax, by Merck Group (2 mentions) β actin ac 15, by Merck Group (2 mentions) Atm mat3 4g10 8, by Merck Group (2 mentions) Phospho chk1 ser317, by R&D Systems (2 mentions) Vinculin, by Merck Group (1 mentions) Dna pkcs, by Fortis Life Sciences (1 mentions) Tcep bond breaker, by Thermo Fisher Scientific (1 mentions) Nupage, by Thermo Fisher Scientific (1 mentions) Anti actin, by Merck Group (1 mentions) Phospho chk2 thr68, by Cell Signaling Technology (1 mentions) Ecl prime, by Cytiva (1 mentions) Parp1, by Cell Signaling Technology (1 mentions) Progesterone receptor a b, by Cell Signaling Technology (1 mentions) Phospho histone h2ax s139, by Cell Signaling Technology (1 mentions) Phospho chk2 t68, by Cell Signaling Technology (1 mentions) Halo software, by Indica Labs (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) β actin, by Merck Group (1 mentions)

6 protocols using phospho atms1981

Western Blot Analysis of DNA Damage Signaling

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Cells were lysed in 1× NuPAGE LDS sample buffer supplemented with 1× Bond-Breaker TCEP (Life Technologies) and 1× Halt protease and phosphatase cocktail (Life Technologies) on ice. The whole-cell lysates were sonicated briefly to break down the chromatin. Samples were boiled for 5 min at 95°C and then loaded to 4%–12% Bris-Tris SDS protein gel or 3%–8% Tris acetate SDS protein gel (Life Technologies).
The antibodies used were as follows: p53 (Calbiochem, DO-1), phospho-p53s15 (Cell Signaling, 9284), p21 (Calbiochem, OP64), DLX2 (Abcam, ab30339), γH2AX (Millipore, 05–636), vinculin (Sigma, V9131), phospho-ATMs1981 (Abcam, ab81292), ATM (Genetex, GTX70103), DNA-PKcs (Bethyl Laboratories, A300-517A), mTOR (Cell Signaling, 4517), ATR (Bethyl Laboratories, A300-137A), SMG1 (Bethyl Laboratories, A300-394A), TTI1 (Bethyl Laboratories, A303-451A), and TEL2 (ProteinTech, 15975-1-AP).

Wang Y., Xu Q., Sack L., Kang C, & Elledge S.J. (2016). A gain-of-function senescence bypass screen identifies the homeobox transcription factor DLX2 as a regulator of ATM–p53 signaling. Genes & Development, 30(3), 293-306.

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Western Blot Analysis of DNA Damage Response

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Cell lysates were obtained using a lithium dodecyl sulfate sample buffer (NuPAGE, Invitrogen). Proteins were resolved on 8 to 15% tris-glycine gels and transferred onto polyvinylidene difluoride membranes. Blots were probed with commercially available primary antibodies and chemiluminescent detection using ECL Prime (Amersham). Antibodies such as Parp1 (#9542), phospho-p53 (S15) (#9284), ATM (#2873), phospho-Chk2 (Thr⁶⁸) (#2197), p21 (#2947), and Puma (#4976) were purchased from Cell Signaling Technology; p53 (#sc-126), Mdm2 (#sc-965), and Wip1 (#sc-20712) from Santa Cruz Biotechnology; γH2A.X (#06-570) from Millipore; and phospho-ATM (S1981) (#ab81292) and phospho-p53 (S46) (#ab76242) from Abcam. Anti-actin (#A5316) from Sigma-Aldrich was used as a loading control.

Yang R., Huang B., Zhu Y., Li Y., Liu F, & Shi J. (2018). Cell type–dependent bimodal p53 activation engenders a dynamic mechanism of chemoresistance. Science Advances, 4(12), eaat5077.

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IHC Analysis of FFPE Tumor Sections

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Five-micron sections of FFPE tissue were mounted on slides, deparaffinized in xylene, and rehydrated in a graded ethanol series. For IHC, antigen retrieval was performed in citrate buffer, pH 6 (VWR), and sections were permeabilized with 0.2% Triton X-100 in PBS. Sections were blocked in 5% goat serum in PBS for 30 min, then incubated overnight in blocking solution with primary antibody at 4°C. Primary antibodies included: phospho-histone H2AX_S139 (Cell Signaling Technology #9718); S9.6 (Millipore Sigma #MABE1095), cleaved caspase 3 (Cell Signaling Technology #9664); phospho-ATM_S1981 (Abcam #81292); phospho-Chk2_T68 (Cell Signaling Technology #2917); progesterone receptor A/B (Cell Signaling Technology #3153). Sections were washed and treated with 0.3% hydrogen peroxide, and signal was developed using the VectaStain Elite ABC-HRP kit with DAB substrate (Vector Laboratories). Sections were counterstained with hematoxylin. IHC staining of PDX tumors was quantified in 3 representative microscopic fields/tumor at 200x magnification using Halo software (Indica Labs).

Cooper H., Smaje C, & Arber S. (1998). Use of health services by children and young people according to ethnicity and social class: secondary analysis of a national survey. BMJ : British Medical Journal, 317(7165), 1047-1051.

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Examining DNA Damage Signaling in A549 Cells

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Example 7

A549 cells were treated with XJB-lapachone at the concentrations indicated in FIG. 5. Western blot was performed 4 4 after treatment to examine the activation of DNA damage signaling. Antibodies used in the Western Blot were β-actin (AC-15) and ATM (MAT3-4G10/8) (from Sigma); Chk1 (2G1D5), phospho-Chk2 (Thr68) (C13C1), Chk2 (1C12) (from Cell Signaling Technology); phospho-Chk1 (Ser317) (from R&D systems); and phospho-ATM (S1981) (from Epitomics). FIG. 9 shows that XJB-lapachone induces DNA damage.

US10968196B2. Substituted β-lapachones for treating cancer (2021-04-06). University of Pittsburgh—Of the Commonwealth System of Higher Education [US]. Inventors: Peter Wipf [US], Chaemin Lim [US], Wei Qian [US], Bennett Van Houten [US].

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XJB-lapachone Induces DNA Damage

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Example 7

US10450289B2. Substituted β-lapachones for treating cancer (2019-10-22). University of Pittsburgh—Of the Commonwealth System of Higher Education [US]. Inventors: Peter Wipf [US], Bennett Van Houten [US], Wei Qian [US], Chaemin Lim [US].

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Western Blot Analysis of DNA Damage Signaling

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Western blot was performed as we previously described (51 (link)). Primary antibodies against ATM and β-actin were obtained from Sigma-Aldrich. Drp1 was from BD Biosciences. Phospho-Drp1 (Ser616), phospho-Chk2 (Thr68), and Chk2 were from Cell Signaling Technology. Phospho-ATM (S1981) was from Epitomics. Complex I subunit NDUFS3, Complex I subunit NDUFB6, and Complex IV subunit II were from abcam.

Qian W., Kumar N., Roginskaya V., Fouquerel E., Opresko P.L., Shiva S., Watkins S.C., Kolodieznyi D., Bruchez M.P, & Van Houten B. (2019). Chemoptogenetic damage to mitochondria causes rapid telomere dysfunction. Proceedings of the National Academy of Sciences of the United States of America, 116(37), 18435-18444.

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