Chromatin Immunoprecipitation:Article Title: Intrinsic signaling pathways modulate targeted protein degradation
Article Snippet: For immunoprecipitation, the anti-FLAG (20 uL agarose/sample, Sigma-Aldrich, #A2220, clone M2) or anti-BRD4 (1 ug/sample, Abcam, #ab128874)antibody was used. .. For Chromatin IP-seq (ChIP-seq), anti-BRD4 antibody (Cell Signaling Technology, #13440, clone E2A7X, rabbit monoclonal, 10 μL) and Spike-in antibody (Active Motif, #61686, 2 μL) were used. anti-BRD4 (1:1000, Cell Signaling Technology, #13440, clone E2A7X), anti-BRD2 (1:1000, Cell Signaling Technology, #5848, clone D89B4), anti-PAR (1:1000, Cell Signaling Technology, #83732, clone E6F6A), anti-PARG (1:1000, Cell Signaling Technology, #66564, clone D4E6X), anti-MEK1 (1:1000, Cell Signaling Technology, #9124), anti-MEK2 (1:1000, Cell Signaling Technology, #9125), anti-ETS1 (Cell Signaling Technology, #14069), anti-PERK (Cell Signaling Technology, #3192), and anti-Cleaved PARP1 (Cell Signaling Technology, #5625), anti-ETS1 (1:1000, Cell Signaling Technology, #14069, clone D8O8A), anti-PERK (1:1000, Cell Signaling Technology, #3192, clone C33E10), and anti-Cleaved PARP1 (1:1000, Cell Signaling Technology, #5625, clone Asp214, D64E10), anti-CDK9(1:1000, Cell Signaling Technology, #2316, clone C12F7), anti-c-MET(1:1000, Cell Signaling Technology, #4560), anti-PSMD4(1:1000, Cell Signaling Technology, #3846), anti-PSMB5(1:1000, Cell Signaling Technology, #12919, clone D1H6B), anti-Rad23B(1:1000, Cell Signaling Technology, #13525, clone D4W7F) were validated by western blotting of human cell lines in the manufacturer's web site (https:// www.cellsignal.com/). anti-BRD3 (1:200, Santa Cruz Biotechnology, sc-515666, clone B-12), anti-CUL2 (1:500, Santa Cruz Biotechnology, sc-166506, clone C-4), anti-b-Actin (1:500, Santa Cruz Biotechnology, sc-47778, clone C4), anti-ER-alpha (1:1000, Santa Cruz Biotechnology, sc-543, clone HC-20), anti-Ub (1:1000, Santa Cruz Biotechnology, sc-8017, clone P4D1), anti-HSP90 (1:500, Santa Cruz Biotechnology, sc-69703, clone 4F10) were validated by western blotting of human cell lines in the manufacturer's web site (https://www.scbt.com/ ja/home). .. Anti-FLAG (Sigma-Aldrich, #A2220, clone M2) was validated in the manufacturer’s web site (https://www.sigmaaldrich.com/JP/ja). anti-TRIP12 (Proteintech, #25303-1-AP) was validated by western blotting of human cell lines in the manufacturer's web site (https:// www.ptglab.co.jp/). anti-VHL (Novus Biologicals, #091504) was validated by western blotting of human cell lines in the manufacturer's web site (https:// www.novusbio.com/japan). anti-BRD4 (Cell Signaling Technology, #13440) was validated by ChIP-seq of human chromatin in the manufacture's web site (https:// www.cellsignal.jp/products/primary-antibodies/brd4-e2a7x-rabbit-mab/13440).
Western Blot:Article Title: Intrinsic signaling pathways modulate targeted protein degradation
Article Snippet: For immunoprecipitation, the anti-FLAG (20 uL agarose/sample, Sigma-Aldrich, #A2220, clone M2) or anti-BRD4 (1 ug/sample, Abcam, #ab128874)antibody was used. .. For Chromatin IP-seq (ChIP-seq), anti-BRD4 antibody (Cell Signaling Technology, #13440, clone E2A7X, rabbit monoclonal, 10 μL) and Spike-in antibody (Active Motif, #61686, 2 μL) were used. anti-BRD4 (1:1000, Cell Signaling Technology, #13440, clone E2A7X), anti-BRD2 (1:1000, Cell Signaling Technology, #5848, clone D89B4), anti-PAR (1:1000, Cell Signaling Technology, #83732, clone E6F6A), anti-PARG (1:1000, Cell Signaling Technology, #66564, clone D4E6X), anti-MEK1 (1:1000, Cell Signaling Technology, #9124), anti-MEK2 (1:1000, Cell Signaling Technology, #9125), anti-ETS1 (Cell Signaling Technology, #14069), anti-PERK (Cell Signaling Technology, #3192), and anti-Cleaved PARP1 (Cell Signaling Technology, #5625), anti-ETS1 (1:1000, Cell Signaling Technology, #14069, clone D8O8A), anti-PERK (1:1000, Cell Signaling Technology, #3192, clone C33E10), and anti-Cleaved PARP1 (1:1000, Cell Signaling Technology, #5625, clone Asp214, D64E10), anti-CDK9(1:1000, Cell Signaling Technology, #2316, clone C12F7), anti-c-MET(1:1000, Cell Signaling Technology, #4560), anti-PSMD4(1:1000, Cell Signaling Technology, #3846), anti-PSMB5(1:1000, Cell Signaling Technology, #12919, clone D1H6B), anti-Rad23B(1:1000, Cell Signaling Technology, #13525, clone D4W7F) were validated by western blotting of human cell lines in the manufacturer's web site (https:// www.cellsignal.com/). anti-BRD3 (1:200, Santa Cruz Biotechnology, sc-515666, clone B-12), anti-CUL2 (1:500, Santa Cruz Biotechnology, sc-166506, clone C-4), anti-b-Actin (1:500, Santa Cruz Biotechnology, sc-47778, clone C4), anti-ER-alpha (1:1000, Santa Cruz Biotechnology, sc-543, clone HC-20), anti-Ub (1:1000, Santa Cruz Biotechnology, sc-8017, clone P4D1), anti-HSP90 (1:500, Santa Cruz Biotechnology, sc-69703, clone 4F10) were validated by western blotting of human cell lines in the manufacturer's web site (https://www.scbt.com/ ja/home). .. Anti-FLAG (Sigma-Aldrich, #A2220, clone M2) was validated in the manufacturer’s web site (https://www.sigmaaldrich.com/JP/ja). anti-TRIP12 (Proteintech, #25303-1-AP) was validated by western blotting of human cell lines in the manufacturer's web site (https:// www.ptglab.co.jp/). anti-VHL (Novus Biologicals, #091504) was validated by western blotting of human cell lines in the manufacturer's web site (https:// www.novusbio.com/japan). anti-BRD4 (Cell Signaling Technology, #13440) was validated by ChIP-seq of human chromatin in the manufacture's web site (https:// www.cellsignal.jp/products/primary-antibodies/brd4-e2a7x-rabbit-mab/13440).
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