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2 protocols using mouse anti igg

1

Chromatin Immunoprecipitation of EBNA1

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ChIP assays were performed as previously described (Chau and Lieberman, 2004 (link)). Briefly 293T cells were plated and transfected as in the plasmid replication assay with FLAG-EBNA1 WT or the Y518F point mutant. Seven days post transfection, cells were synchronized with aphidicolin and S phase cells were harvested for either native or crosslinked ChIP. TaqMan qPCR performed using primers and probe detecting EBV B95.8 genome coordinates 8401–8521 adjacent to FR was designed by Themo-Fisher. Antibodies used were as follows: mouse anti-IgG (Santa Cruz Biotechnology), Trimethyl-histone H3 (Lys4) (Millipore, cat no. 07–473), anti-flag M2 mouse monoclonal (Sigma Aldrich, cat no. F1804).
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2

Immunoprecipitation and Western Blot Analysis

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Rabbit-anti-ID3 (Cell Signaling, Cat#9837) and for IP of endogenous ID3 we used agarose beads conjugated with mouse-anti-ID3(Santa Cruz, Cat#sc-56712). Mouse-anti-beta-Actin (Santa Cruz, Cat#sc-47778). Mouse-anti-Vinculin (Santa Cruz, Cat#sc-25336). Mouse-anti-phospho-Histone H2A.X (S139) (Merck, Cat#05-636). Rabbit-anti-phospho-Histone H2A.X (S139) (Abcam, Cat# ab2893). Rabbit-anti-RAD51 (Calbiochem, Cat#PC130). Rabbit-anti-RAD51 (Abcam, Cat#ab176458). Rabbit-anti-XRCC4 (GenTex, Cat#GTX109632). Rabbit-anti-phospho-RPA (S4/S8) (Bethyl, Cat#A300-245A). Mouse-anti-Histone H2B (Abcam, Cat#ab52484). Rabbit-anti-DNA-PKsc (Cell Signaling, Cat#4602). Rabbit-anti-CtIP (Abcam, Cat#70163). Goat-anti-MDC1 (Santa Cruz, Cat#sc-27737). Rabbit-anti-MDC1 (Abcam, Cat#ab11169). Rabbit-anti-NBS1 (Novus Biologicals, Cat#NB100-143). Mouse-anti-RAD50 (Abcam, Cat#ab89). Rabbit-anti-RECQL (Abcam, Cat#ab151501). Mouse-anti-RPA32 (Santa Cruz, Cat#sc-53496). Mouse-anti-Flag-HRP (Sigma, Cat#A8592). Mouse-anti-IgG (Santa Cruz, Cat#sc-2025). Mouse-anti-CenpF (BD bioscience, Cat#610768). Goat-anti-mouse IgG-HRP (Cell Signaling, Cat#7076P2). Goat-anti-rabbit IgG-HRP (Cell Signaling, Cat#7074S). Goat-anti-mouse IgG-AlexaFluor 594 (Molecular Probes, Cat#A11005). Goat-anti-rabbit IgG-AlexaFluor 488 (Molecular Probes, Cat#A11008).
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