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2 protocols using parp 1 sc 53643

1

Antibody Panel for Immunoblotting DNA Repair Proteins

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The following antibodies for immunoblotting were used. APE1, Pol β and XRCC1 antibodies were kindly provided by Dr G. Dianov. PNKP antibodies were also kindly provided by Prof M. Weinfeld. ATM (sc-23921), DNA-Pk (sc-9051), Ku-86 (sc-9034), PARP-1 (sc-53643) and RAD51 (sc-8349) antibodies were from Santa Cruz Biotechnology (Heidelberg, Germany); ATR antibodies (ab2905) were from Abcam (Cambridge, UK); 53BP1 (A300-272A) antibodies were from Bethyl Laboratories (Montgomery, USA); and actin (A5441) antibodies were from Sigma-Aldrich (Gillingham, UK).
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2

Investigating Cellular Pathways through siRNA

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The DUB siRNA library (ON-TARGETplus), USP9X siRNA pool and individual siRNA targeting USP9X (USP9X_6 5′-AGAAAUCGCUGGUAUAAAU-3′ and USP9X_8 5′-GUACGACGAUGUAUUCUCA-3′) were from Horizon Discovery (Cambridge, UK). The non-targeting control siRNA (AllStars Negative Control siRNA) was from Qiagen (Manchester, UK). The following antibodies were used: PARP-1 (sc-53643) and Mcl-1 (sc-819; both Santa Cruz Biotechnology, Heidelberg, Germany), γH2AX (05-636; Merck-Millipore, Watford, UK), USP9X (A301-350A) and 53BP1 (A300-272A; both Bethyl Labs, Montgomery, AL), LC3B (2775) and β-Galactosidase (9860; both Cell Signaling Technology, London, UK), Pericentrin (ab4448; Abcam, Cambridge, UK), CEP55 (23891-1-AP) and CEP131 (25735-1-AP; both Proteintech, Manchester, UK) and tubulin (T6199; Sigma-Aldrich, Gillingham, UK). Goat anti-mouse Alexa Fluor 555 (A21422) or goat anti-rabbit Alexa Fluor 488 (A11008) secondary antibodies for immunofluorescence were from Life Technologies (Paisley, UK).
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