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Patm s1981

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The PATM S1981 is a laboratory equipment product designed for specific applications. It is a device that performs a core function, but a detailed description while maintaining an unbiased and factual approach is not available at this time.

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Lab products found in correlation

Pchk2 t68, by Cell Signaling Technology (6 mentions) P chk1, by Cell Signaling Technology (3 mentions) γh2ax, by Merck Group (3 mentions) P p53 s15, by Cell Signaling Technology (3 mentions) P atm s1981, by Cell Signaling Technology (2 mentions) P tbk1 s172, by Cell Signaling Technology (2 mentions) P histone h3 s10, by Cell Signaling Technology (2 mentions) P histone h3 t11, by Cell Signaling Technology (2 mentions) Coxii, by Abcam (2 mentions) Parkin, by Santa Cruz Biotechnology (2 mentions) Gapdh and actin, by Merck Group (2 mentions) Secondary hrp linked antibodies, by GE Healthcare (2 mentions) Tom20, by Santa Cruz Biotechnology (2 mentions) Pink1, by Cell Signaling Technology (2 mentions) Ha 11, by Fortrea (2 mentions) Chemidoc imaging system, by Bio-Rad (2 mentions) Tim23, by BD (2 mentions) γh2ax, by Santa Cruz Biotechnology (2 mentions) Lamin b1, by Abcam (2 mentions) Gapdh, by Santa Cruz Biotechnology (2 mentions)

20 protocols using patm s1981

1

Western Blot Analysis of Cellular Proteins

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Cells were lysed using 2x LDS buffer containing 50mM DTT and boiled for 15 min. Approximately 50 μg of protein lysate was loaded onto 4%–12% Bis-Tris gels. Proteins were transferred onto PVDF membranes and blocked with 5% non-fat powdered milk. Primary antibodies were incubated 4°C overnight. Antibodies used: PINK1, p-ATM S1981, LC3B, TBK1, p-TBK1 S172, p-Histone H3 S10 and p-Histone H3 T11 (Cell Signaling), pATM S1981, COXII (Abcam), ATM (Cell Signaling or Sigma), Tim23 (BD Biosciences), Mfn2 (in-house), TOM20, Parkin, p53 (Santa Cruz), GAPDH and actin (Sigma), p62 (Cedarlane), and HA.11 (Covance). Secondary HRP-linked antibodies (GE Healthcare) were incubated at RT for 1 hr. Blots were exposed using peroxidase-based ECL (Pierce) detected by a ChemiDoc Imaging System (BioRad).
Sarraf S.A., Sideris D.P., Giagtzoglou N., Ni L., Kankel M.W., Sen A., Bochicchio L.E., Huang C.H., Nussenzweig S.C., Worley S.H., Morton P.D., Artavanis-Tsakonas S., Youle R.J, & Pickrell A.M. (2019). PINK1/Parkin Influences Cell Cycle by Sequestering TBK1 at Damaged Mitochondria, Inhibiting Mitosis. Cell reports, 29(1), 225-235.e5.
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2

Western Blot Analysis of Cellular Proteins

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Cells were lysed using 2x LDS buffer containing 50mM DTT and boiled for 15 min. Approximately 50 μg of protein lysate was loaded onto 4%–12% Bis-Tris gels. Proteins were transferred onto PVDF membranes and blocked with 5% non-fat powdered milk. Primary antibodies were incubated 4°C overnight. Antibodies used: PINK1, p-ATM S1981, LC3B, TBK1, p-TBK1 S172, p-Histone H3 S10 and p-Histone H3 T11 (Cell Signaling), pATM S1981, COXII (Abcam), ATM (Cell Signaling or Sigma), Tim23 (BD Biosciences), Mfn2 (in-house), TOM20, Parkin, p53 (Santa Cruz), GAPDH and actin (Sigma), p62 (Cedarlane), and HA.11 (Covance). Secondary HRP-linked antibodies (GE Healthcare) were incubated at RT for 1 hr. Blots were exposed using peroxidase-based ECL (Pierce) detected by a ChemiDoc Imaging System (BioRad).
Sarraf S.A., Sideris D.P., Giagtzoglou N., Ni L., Kankel M.W., Sen A., Bochicchio L.E., Huang C.H., Nussenzweig S.C., Worley S.H., Morton P.D., Artavanis-Tsakonas S., Youle R.J, & Pickrell A.M. (2019). PINK1/Parkin Influences Cell Cycle by Sequestering TBK1 at Damaged Mitochondria, Inhibiting Mitosis. Cell reports, 29(1), 225-235.e5.
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3

Immunoblotting Antibody Panel for DNA Damage

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For immunoblotting experiments, we utilized the following antibodies: −γH2AX(S139) (Cell Signaling #80312), H2AX (Sigma‐Aldrich # 07627), pRPA32(T21) (Abcam #ab109394), RPA32 (Cell Signaling #35869), −pATR(T1986) (Cell Signaling #58014 and #30632), ATR (Cell Signaling #13934), pCHK1(S345) (Cell Signaling #12302), CHK1 (Cell Signaling #2360), pATM(S1981) (Abcam #ab81292), ATM (Cell Signaling #2873), pCHK2(T68) (Cell Signaling #2197), CHK2 (Cell Signaling #6334), pDNA‐PK(S2056) (Abcam# ab18192), DNA‐PK (Abcam# ab32566), vinculin (Sigma‐Aldrich # V9131), histone H3 (Sigma‐Aldrich #07‐690), and SLFN11 (Santa‐Cruz #sc‐374339). All antibodies were diluted at 1:1,000 except SLFN11 1:2,000, tubulin 1:4,000, and vinculin 1:4,000, and all secondary antibodies were diluted at 1:4,000.
Schultz C.W., Zhang Y., Elmeskini R., Zimmermann A., Fu H., Murai Y., Wangsa D., Kumar S., Takahashi N., Atkinson D., Saha L.K., Lee C., Elenbaas B., Desai P., Sebastian R., Sharma A.K., Abel M., Schroeder B., Krishnamurthy M., Kumar R., Roper N., Aladjem M., Zenke F.T., Ohler Z.W., Pommier Y, & Thomas A. (2023). ATR inhibition augments the efficacy of lurbinectedin in small‐cell lung cancer. EMBO Molecular Medicine, 15(8), e17313.
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4

Comprehensive DNA Damage Signaling Assay

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GFP (Abcam catalog no. ab6556), TRF1(Abcam catalog no. ab10579), GAPDH (Santa Cruz catalog no. sc-47724), OGG1 (Abcam catalog no. ab124741), Actin Cell Signaling catalog no. 3700), Lamin B1 Abcam catalog no. ab16048), Lamin A/C (Cell Signaling catalog no. 4777), γH2AX (Santa Cruz catalog no. sc-517348), 53BP1 (Novous catalog no. NB100-304), TRF2 (Novous catalog no. NB110-57130), MDM2 (Cell Signaling catalog no. 86934), p53(Santa Cruz catalog no. sc-126), p21 (Cell Signaling catalog no. 2947), p16 (Proteintech catalog no. 10883-1-AP), pRB S807/811 (Cell Signaling catalog no. 8516), pCHK2 T68(Cell Signaling catalog no. 2197), pCHK1 S317 (Cell Signaling catalog no. 12302), pATM S 1981 (Abcam catalog no. ab81292), CHK1 (Cell Signaling catalog no. 2360), H3K27me3 (Cell Signaling catalog no. 9733), H3K27Ac (Cell Signaling catalog no. 8173), LSD1 (Cell Signaling catalog no. 2184), cGAS (Cell Signaling catalog no. 66546), p62 (Cell Signaling catalog no. 39749).
Barnes R.P., de Rosa M., Thosar S.A., Detwiler A.C., Roginskaya V., Van Houten B., Bruchez M.P., Stewart-Ornstein J, & Opresko P.L. (2022). Telomeric 8-oxo-guanine drives rapid premature senescence in the absence of telomere shortening. Nature Structural & Molecular Biology, 29(7), 639-652.
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5

Western Blot and Immunofluorescence Analysis

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Antibodies used for Western Blot were pChk1 S345 (Cell Signaling, CS2348, lot# 18, 1:1000), Chk1 (Santa Cruz, SC8408, lot# A1713, 1:1000), pChk2 T68 (Cell Signaling, CS2661, lot# 13, 1:1000), Chk2 (Cell Signaling, CS3440, lot# 4, 1:1000), γH2AX (Millipore, 05-636, lot# 3824772, 1:1000), p53 (Santa Cruz, SC-126, lot# E2521, 1:2000), p21 (BD Biosciences, 556430, lot# 1173681, 1:1000), ATM (Abcam, ab78, lot# GR3286342-11, 1:2500) pATM S1981 (Abcam, ab81292, 1:2500) tubulin (Millipore, CP06 OS, lot#3239856 1:2000), PRDX2 (Abcam, ab109367, lot# 1000538-1 1:2000) and PRDXSO2/3 (Abcam, ab16830, lot# GR3294252-3, 1:1000).
Antibodies used for IF were LaminB1 (Merck, ZRB1143, lot# Q3250182, 1:500) and p21 (Merck, ZRB1141, lot# 3307439, 1:800).
van Soest D.M., Polderman P.E., den Toom W.T., Keijer J.P., van Roosmalen M.J., Leyten T.M., Lehmann J., Zwakenberg S., De Henau S., van Boxtel R., Burgering B.M, & Dansen T.B. (2024). Mitochondrial H2O2 release does not directly cause damage to chromosomal DNA. Nature Communications, 15, 2725.
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6

Western Blot Analysis of Akt and ATM Signaling

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Following treatment, cells were lysed in ice cold radioimmunoprecipitation assay (RIPA) buffer supplemented with protease inhibitor cocktail and PMSF (AppliChem, Darmstadt, Germany). Lysates were boiled in 1× SDS-PAGE sample buffer (25 mM Tris-HCl, pH 6.8; 10% glycerol, 2% SDS, 2.5% β-mercaptoethanol, 0.005% bromophenol blue) and equivalent amounts of protein were electrophoresed on SDS-PAGE gels. PVDF membranes were blotted with antibodies that recognize Akt, (#2938, 1:1000), p-Akt Thr308 (# 2965, 1:2000), p-Akt Ser473 (# 4060, 1:1500), GAPDH (#2118, 1:1000), (all Cell Signaling, Frankfurt, Germany), ATM (# 1549-1), p-ATM S1981 (# 2152-1, Abcam, Cambridge, UK) and subsequently incubated with anti-rabbit/anti-mouse HRP (horseradish peroxidase)-linked IgG antibodies (1:5000, Millipore, Darmstadt, Germany). Proteins were visualized using enhanced chemiluminescence detection system (Amersham, Munich, Germany). Generally, specific expression was normalized to the mean value of the total blot, or, when specified, to the untreated control. Raw data of Western Blots is provided in Figure S7.
Schötz U., Balzer V., Brandt F.W., Ziemann F., Subtil F.S., Rieckmann T., Köcher S., Engenhart-Cabillic R., Dikomey E., Wittig A, & Arenz A. (2020). Dual PI3K/mTOR Inhibitor NVP-BEZ235 Enhances Radiosensitivity of Head and Neck Squamous Cell Carcinoma (HNSCC) Cell Lines Due to Suppressed Double-Strand Break (DSB) Repair by Non-Homologous End Joining. Cancers, 12(2), 467.
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7

Antibody Panel for DNA Damage Response

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Primary antibodies used in this study were Flag M2 (Sigma, F1804), Myc (Santa Cruz Biotechnology, SC-40), H2AX (Millipore, 07-627), γH2AX (Millipore, 05-636), H2AZ (Cell Signaling, 2718S), macroH2A (Abcam, AB37264), H2A.Bbd (Millipore, 06-1319), ZMYM3 (Abcam, AB106626), ATM (Santa Cruz Biotechnology, SC-135663), pATM S1981 (Abcam, AB81292), β-tubulin (Abcam, AB6046), RAD51 (Abcam, AB88572), RAP80 (Bethyl Laboratories, A300-763A), ABRA1 (Abcam, AB139191), BRCA1 (Santa Cruz Biotechnology, SC-6954), HRP-linked anti-GST (Sigma, A7340), MBP (Abcam, AB9084), phospho-H3 S10 (Cell Signaling, 3377), 53BP1 (Novus Biologicals, NB100-304), RAD18 (Cell Signaling, 9040), RPA2 (Abcam, AB2175), pRPA2 S33 (Bethyl Laboratories, A300-246), pRPA2 S4/S8 (Bethyl Laboratories, A300-245), Chk1 (Santa Cruz Biotechnology, SC-8408), and pChk1 (Cell Signaling, 2348). For Western blotting analysis, secondary antibodies HRP-linked anti-rabbit IgG and HRP-linked anti-mouse IgG were purchased from Cell Signaling. For immunofluorescence, Alexa fluor 488 goat anti-rabbit and Alexa fluor 594 goat anti-mouse were used (Invitrogen).
Leung J.W., Makharashvili N., Agarwal P., Chiu L.Y., Pourpre R., Cammarata M.B., Cannon J.R., Sherker A., Durocher D., Brodbelt J.S., Paull T.T, & Miller K.M. (2017). ZMYM3 regulates BRCA1 localization at damaged chromatin to promote DNA repair. Genes & Development, 31(3), 260-274.
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8

Innate Immunity and DNA Damage Response Pathway Analysis

Cited 1 time
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Etoposide, 4-hydroxytamoxifen, puromycin, and polybrene were purchased from Sigma. Hygromycin was from Gemini Bio Products, and luciferin was from PerkinElmer. Recombinant human IL1α protein (used at 20 ng/ml) was from R&D Systems, and p38 MAPK inhibitor from Selleck Chemicals.
The following antibodies were described previously: Flag, p16, Lamin B1, GAPDH, and GFP5 (link). Other antibodies used include: γH2AX (Abcam #ab2893 and Cell Signaling Technology #9718), STING (Cell Signaling Technology #13647 and LSBio #LS-B7237), cGAS (Cell Signaling Technology #15102 and Santa Cruz Biotechnology #sc-245858), human IL1α (Abcam #ab9614), IL8 (Abcam #ab18672), p-ATM S1981 (Abcam #ab81292), p-p53 S15 (Cell Signaling Technology #9284), p-p65 S536 (Cell Signaling Technology #3033), p65 (Cell Signaling Technology 8242), NRas (Santa Cruz Biotechnology #sc-31), CD3 (Abcam #ab16669), Mac2 (BioLegend #125401), mouse IL1α (R&D Systems #AF-400), p21 (#sc-471), and H3 (Active Motif #39763).
Dou Z., Ghosh K., Vizioli M.G., Zhu J., Sen P., Wangensteen K.J., Simithy J., Lan Y., Lin Y., Zhou Z., Capell B.C., Xu C., Xu M., Kieckhaefer J.E., Jiang T., Shoshkes-Carmel M., Ahasan Al Tanim K.M., Barber G.N., Seykora J.T., Millar S.E., Kaestner K.H., Garcia B.A., Adams P.D, & Berger S.L. (2017). Cytoplasmic chromatin triggers inflammation in senescence and cancer. Nature, 550(7676), 402-406.
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9

Comprehensive DNA Damage Response Analysis

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GAPDH (Santa Cruz sc-47724; 1:5000 WB), α-tubulin (Millipore 05-829; 1:5000 WB), OGG1 (Abcam ab124741; 1:500), MUTYH (Abnova H00004595-M01; 1:500 WB); γH2AX (Santa Cruz sc-517348; 1:200 IF), 53BP1 (Novus NB100-304; 1:1000 IF), p53 (Santa Cruz sc-126; 1:200 WB), p21 (Cell Signaling 2947; 1:1000 IF and 1:2000 WB), pCHK2 T68 (Cell Signaling 2197; 1:1000 WB), pCHK1 S345 (Cell Signaling 2348; 1:500 WB), pATM S 1981 (Abcam ab81292; 1:2000 WB), Chk2 (Cell Signaling 3440; 1:1000 WB), CHK1 (Cell Signaling 2360; 1:1000 WB), ATM (Sigma A1106; 1:500 WB); PAR (10H) (Enzo ALX-804-220-R100, 1:500 WB, 1:100 IF), APE1 (Cell Signaling 10519S; 1:1000 WB); Cyclin A (Santa Cruz sc-271682; 1:500 WB).
De Rosa M., Barnes R.P., Nyalapatla P.R., Wipf P, & Opresko P.L. (2023). OGG1 and MUTYH repair activities promote telomeric 8-oxoguanine induced cellular senescence. bioRxiv.
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10

Innate Immunity and DNA Damage Response Pathway Analysis

Cited 1 time
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Etoposide, 4-hydroxytamoxifen, puromycin, and polybrene were purchased from Sigma. Hygromycin was from Gemini Bio Products, and luciferin was from PerkinElmer. Recombinant human IL1α protein (used at 20 ng/ml) was from R&D Systems, and p38 MAPK inhibitor from Selleck Chemicals.
The following antibodies were described previously: Flag, p16, Lamin B1, GAPDH, and GFP5 (link). Other antibodies used include: γH2AX (Abcam #ab2893 and Cell Signaling Technology #9718), STING (Cell Signaling Technology #13647 and LSBio #LS-B7237), cGAS (Cell Signaling Technology #15102 and Santa Cruz Biotechnology #sc-245858), human IL1α (Abcam #ab9614), IL8 (Abcam #ab18672), p-ATM S1981 (Abcam #ab81292), p-p53 S15 (Cell Signaling Technology #9284), p-p65 S536 (Cell Signaling Technology #3033), p65 (Cell Signaling Technology 8242), NRas (Santa Cruz Biotechnology #sc-31), CD3 (Abcam #ab16669), Mac2 (BioLegend #125401), mouse IL1α (R&D Systems #AF-400), p21 (#sc-471), and H3 (Active Motif #39763).
Dou Z., Ghosh K., Vizioli M.G., Zhu J., Sen P., Wangensteen K.J., Simithy J., Lan Y., Lin Y., Zhou Z., Capell B.C., Xu C., Xu M., Kieckhaefer J.E., Jiang T., Shoshkes-Carmel M., Ahasan Al Tanim K.M., Barber G.N., Seykora J.T., Millar S.E., Kaestner K.H., Garcia B.A., Adams P.D, & Berger S.L. (2017). Cytoplasmic chromatin triggers inflammation in senescence and cancer. Nature, 550(7676), 402-406.
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