Dnapkcs ps2056

Manufactured by Abcam

DNAPKcs-pS2056 is a monoclonal antibody that recognizes the phosphorylated form of the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) at serine 2056. DNA-PKcs is a key component of the non-homologous end joining (NHEJ) pathway, which is responsible for the repair of double-strand breaks in DNA.

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Lab products found in correlation

Amersham imager 600, by GE Healthcare (1 mentions) Ibind western system, by Thermo Fisher Scientific (1 mentions) Topoisomerase 1, by BD (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Ctdsp1, by Abcam (1 mentions) Amersham imager, by Cytiva (1 mentions) γh2ax, by Merck Group (1 mentions) Chk1 ps345, by Cell Signaling Technology (1 mentions) Anti flag antibody, by Merck Group (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Dna pkcs, by Cell Signaling Technology (1 mentions) Myc sc 40, by Santa Cruz Biotechnology (1 mentions) Rpa32 ps33, by Fortis Life Sciences (1 mentions) Rpa32 pt21, by Abcam (1 mentions) Rpa32 ps4 8, by Fortis Life Sciences (1 mentions) Rfwd3, by Fortis Life Sciences (1 mentions)

3 protocols using dnapkcs ps2056

Western Blot Protein Analysis Protocol

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Cells cultured in 6-well plates were scraped into an ice-cold RIPA buffer. Samples were clarified by centrifugation at 15000 rpm for 15 min at 4°C. We used the iBind Western System (Invitrogen) and performed the imaging using the Amersham Imager 600 instrument (GE Healthcare, Little Chalfont, UK). Primary antibodies included antibodies against β-actin (Cell Signaling Technology), CTDSP1 (Abcam), Topoisomerase I (BD Pharmingen), and DNAPKcs-pS2056 (Abcam). Antigen/antibody complexes were visualized by enhanced chemiluminescence (ECL detection system).

Matsuoka H., Ando K., Swayze E.J., Unan E.C., Mathew J., Hu Q., Tsuda Y., Nakashima Y., Saeki H., Oki E., Bharti A.K, & Mori M. (2020). CTDSP1 inhibitor rabeprazole regulates DNA-PKcs dependent topoisomerase I degradation and irinotecan drug resistance in colorectal cancer. PLoS ONE, 15(8), e0228002.

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Immunofluorescence Analysis of DNA Damage Markers

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For immunofluorescence analysis, cells were cultured on 35 mm² dishes with coverslips, and treated with BPQDs and indicated dose of irradiation. Cells were fixed with 4% paraformaldehyde, and permeabilized in 0.5% Triton X-100 for additional 20 min, and blocked in 5% bovine serum albumin. The samples were incubated with the following antibodies: γH2AX (Millipore, 05-636, 1:1000), 53BP1 (Novusbio, NB100-904, 1:200), DNA-PKcs-ps2056 (Abcam, ab18192, 1:1000), Alexa-488-conjugated anti-rabbit secondary antibodies (CST, 4916, 1:1000), and Alexa-488-conjugated anti-mouse secondary antibodies (CST, 4408, 1:1000).

Lang Y., Tian X., Dong H.Y., Zhang X.X., Yu L., Li M., Gu M.M., Gao D, & Shang Z.F. (2022). Black Phosphorus Quantum Dots Enhance the Radiosensitivity of Human Renal Cell Carcinoma Cells through Inhibition of DNA-PKcs Kinase. Cells, 11(10), 1651.

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DNA Damage Response Antibodies

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The following antibodies were used in this study: PRP19 (ab27692, 1:400 IF, 1:1000 WB), ATM pS1981 (ab81292, 1:1000 WB), DNAPKcs pS2056 (ab18192, 1:1000 WB) and RPA32 pT21 (ab109394, 1:5000 WB) antibodies were from Abcam, CHK1 (SC-8404, 1:1000 WB), Myc (SC-40, 1:1000 WB) and HA (SC-7392, 1:1000 WB) antibodies were from Santa Cruz; RPA32 antibodies (MA1–26418, 1:500 IF, 1:1000 WB) were from Thermo, ATM (A300–299A, 1:1000 WB), RPA32 pS4/8 (A300–245A, 1:1000 WB), RPA32 pS33 (A300–246A, 1:5000 WB), RFWD3 (A301–397A, 1:250 IF, 1:1000 WB) antibodies were from Bethyl; ubiquitin antibodies were from Covance (MMS-257P, 1:1000 WB); CDC5L antibodies were from BD-Bioscience (612362, 1:1000 WB); anti-FLAG antibodies were from Sigma (F1804, 1:800 IF, 1:1000 WB); CHK1 pS345 (2348, 1:1000 WB), GAPDH (5174, 1:1000 WB), DNAPKcs (12311, 1:1000 WB) and tubulin (2144, 1:1000 WB) antibodies were from Cell Signaling.

Dubois J.C., Yates M., Gaudreau-Lapierre A., Clément G., Cappadocia L., Gaudreau L., Zou L, & Maréchal A. (2017). A phosphorylation-and-ubiquitylation circuitry driving ATR activation and homologous recombination. Nucleic Acids Research, 45(15), 8859-8872.

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