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3 protocols using flag hrp a8592

1

Antibody and Reagent Protocols for Cell Death Analysis

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Antibodies for PARP (9532, 1:1000), ULK1 (8054, 1:1000), p62 (5114, 1:1000), AIF (5318, 1:1000) and Lamin B (12586, 1:1000) were purchased from Cell Signaling Technology (MA, USA). TFG (ab156866, 1:10,000) and GSDMD (ab210070, 1:1000) were from Abcam. Caspase-1 (AG-20B-0048-C100, 1:1000) were purchased from AdipoGen (CA, USA). HA (12CA5, 1:1000 for IP) and HA-HRP (3F10, 1:5000) were from Roche Life Science (Switzerland). Flag (F3165, 1:500 for IP) and Flag-HRP (A8592, 1:5000), LC3B (L7543, 1:1000) and β-actin (A2228, 1:10,000) were from Sigma–Aldrich (MO, USA). Ub (P4D1, 1:1000), TRAF3 (H-122, 1:1000), Tubulin (SC-8035, 1:1000), and rabbit IgG (sc-2027, 1:1000) were from Santa Cruz Biotechnology (CA, USA). COX4 (ARG66326, 1:1000) were purchased from Arigo Biolaboratories. V5 (R96025, 1:500 for IP) and V5-HRP (R96125, 1:5000) were from Thermo Fisher Scientific (MA, USA).
MitoSox™ Red Mitochondrial Superoxide Indicator (M36008), SYTOX™ Blue Dead Cell Stain (S34857), and SYTOX™ Red Dead Cell Stain (S34859) were purchased from Thermo Fisher Scientific (MA, USA). LPS, Nigericin, Z-VAD-FMK (Z-VAD), Z-Leu-Leu-Leu-al (MG132) were purchased from Sigma–Aldrich (MO, USA). Necrostatin-1 (NEC-1) was purchased from ApexBio (Texas, USA). Cycloheximide (CHX), E-64d, and Pepstatin A were purchased from Cayman Chemical (Michigan, USA).
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2

Comprehensive Antibody Panel for Chromatin Studies

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Antibodies recognizing SMC2 (A300-058A), SMC4 (A300-064A), NCAPH2 (A302-275A), NCAPD3 (A300-604A), NCAPG2 (A300-605A), TFIIIC-220 (A301-291A), TFIIIC-102 (A301-238A), TFIIIC-63 (A301-242A), SET1 (A300-290A), and NPAT (A302-772A) were purchased from Bethyl Laboratories. Anti-SMC3 antibody (sc-292645) was purchased from Santa Cruz Biotechnology. Anti-RAD21 antibody (05-908) was purchased from Millipore. Antibodies recognizing H3K4me3 (39915), H3K27ac (39133), and WDR5 (61485) were purchased from Active Motif. Antibodies recognizing CTCF (ab70303) and H3 (ab1791) were purchased from Abcam. Flag-HRP (A8592) and α-tubulin (T6199) antibodies were purchased from Sigma-Aldrich.
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3

Flag-M2 Affinity Purification Protocol

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Flag-M2 affinity agarose gel was from Raygene; Mono Q and Superose 6 were from GE Healthcare; polyethylenimine (PEI) was from Polysciences (23966); HEK293A and Expi293F cells were from Invitrogen Inc.; and culture medium was from Sino Biological Inc. Antibodies against phosphorylated-S6K (Thr 389) were from Cell Signaling Technology; Flag-HRP (A8592) was from Sigma; and Horseradish peroxidase-labeled anti-mouse and anti-rabbit secondary antibodies were from AbMart.
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