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3 protocols using abs130593

1

Western Blot Antibody Validation Protocol

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Antibodies against H3 (1: 5000; ab1791), H3pT11 (1:3000; ab5168) and SETX (1:3000; ab243904) were purchased from Abcam; antibody against FLAG M2 (1:3000; F1804-1MG) was obtained from Sigma-Aldrich; antibodies against Sir2 (1:500; sc-6667) and LC3 (1:1000; sc-398822) were purchased from Santa Cruz Biotechnology; antibodies against Myc (1:5000; 60003-2-1 g), GAPDH (1:10000; 10494-1-AP), beta-actin (1:10000; 20536-1-AP), GFP (1:5000; 66002-1-Ig) and goat polyclonal anti-mouse IgG (1:5000; SA00001-1) were obtained from Proteintech; antibody against Rif1 (1:3000; 95558 S) was purchased from Cell Signaling Technology; antibodies against CDKN1A/p21 (1:3000; A11454), and PPP1CA (1:5000; A12468) were purchased from Abclonal; antibody against CBP (1:3000; abs130593) was purchased from Absin. Antibody against Glc7 (1:5000) was custom-made in Abclonal and its specificity was confirmed by dot blots with Glc7 peptide (Supplementary Fig. S11b). H3pT11 was validated for ChIP assay in H3T11A mutant and PKM2 knockdown HUVECs (Supplementary Fig. S11c, d).
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2

Antibody Verification for Histone Modifications

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Antibodies against anti-H3 (1:5000; ab1791), anti-H4 (1:5000; ab10158) and H3pT11 (1:5000; ab5168) were purchased from Abcam; antibodies against H3K79me1 (1: 1000; A2367), H3K79me2 (1: 5000; A2368), H3K79me3 (1:5000; A2369), anti-His (1:5000; AE0039) and anti-FLAG (1:5000; AE024) were purchased from Abclonal; antibody against Sir2 (1:500; sc-6667) was purchased from Santa Cruz Biotechnology; antibodies against GAPDH (1:10,000; 10494-1-AP), GFP (1:5000; 66002-1-1 g), Myc (1:5000; 60003-2-1 g), goat polyclonal anti-mouse IgG (1:5000; SA00001-1) and goat polyclonal anti-rabbit IgG (1:5000; SA00001-2) were obtained from Proteintech; antibody against FLAG M2 (1:3000; F1804-1MG) was obtained from Sigma-Aldrich; antibody against H4K16ac (1:2000; 07-329) was obtained from EMD Millipore; antibody against CBP (1:2000; Abs130593) was purchased from Absin Bioscience Inc. The custom-made antibodies against Acs2, Pyk1, Sam1 and Shm2 were produced by Covance Inc. and their specificity was verified by immunoblots of cell lysates of the corresponding histone point mutants, gene deletion or knockdown mutants31 (link).
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3

Antibody Validation for Histone Modifications

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Antibody against anti-histone H2A.Z (1:5000; ab4626) was purchased from Abcam; antibody against Rpd3 (1:500; sc-514160) was purchased from Santa Cruz Biotechnology; antibodies against GFP (1:5000; 66002-1-1g), Myc (1:5000; 60003-2-1g), goat polyclonal anti-mouse IgG (1:5000; SA00001-1), and goat polyclonal anti-rabbit IgG (1:5000; SA00001-2) were obtained from Proteintech; antibody against pan–acetyl-lysine (1:2000, 9441S) was purchased from Cell Signaling Technology; antibody against FLAG M2 (1:3000; F1804-1MG) was obtained from Sigma-Aldrich; antibody against CBP (1:2000; Abs130593) was purchased from Absin Bioscience Inc.; and antibodies against Ino80 (1:500) and Ino80 K929ac (1:500) were custom-made in Abclonal. The specificity of the custom-made antibodies was confirmed by immunoblot analysis with cell extract or IP of corresponding mutants (Fig. 3D and fig. S9).
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