Anti phospho ripk1

Western blotting experiment was described previously (Zhao et al., 2019 (link)), and the detection of RIPK3 or MLKL oligomers was performed according to the method described by Wang et al. (2019) (link). The following antibodies were used for the experiments: anti-FADD (1:1000, 610399) and anti-RIPK1 (1:3000, 610458) (BD Transduction Laboratories, San Jose, CA); anti-phospho-RIPK3 (1:1000, ab195117), anti-MLKL (1:1000, ab172868), anti-phospho-MLKL (1:1000, ab196436) (Abcam, Cambridge, MA, United States); anti-RIPK1 (1:3000, 3493), anti-RIPK3 (1:3000, 15828), anti-Myc tag (1:2000, 2276), anti-HA tag (1:2000, 3724), anti-Flag tag (1:2000, 14793), anti-TRAF2 (1:1000, 4724), anti-phospho-RIPK1 (1:1500, 31122) and anti-phospho-RIPK3 (1:1000, 57220) [Cell Signaling Technology (CST), Beverly, MA]; anti-TRADD (1:1000, ABP52634) and anti-GAPDH (1:1500, ABP52783) (Abbkine, Redlands, CA, United States); anti-β-actin (1:3000, A5441) (Sigma-Aldrich). All western blot assays were performed three times, and the representative results are shown.

The following antibodies were used in this study: anti-RIPK1 (BD Biosciences, San Jose, CA, USA), anti-phospho-RIPK1 (Ser166; Cell Signaling Technology, Danvers, MA, USA), anti-phospho-hRIPK1 (Ser166 [39 (link)]; Cell Signaling Technology), anti-RIPK3 (Novus Biologicals, Littleton, CO, USA), anti-phospho-hRIPK3 (Ser227) (Cell Signaling Technology), anti-MLKL (Abgent, San Diego, CA, USA), anti-phospho-MLKL (Abcam, Cambridge, UK), anti-β-actin (Sigma-Aldrich, USA), anti-β-tubulin (Cell Signaling Technology), anti-cleaved caspase-3 (Cell Signaling Technology), anti-cleaved PARP (Cell Signaling Technology), anti-phospho-IκBα (Cell Signaling Technology), and anti-IκBα (Santa Cruz Biotechnology).

The following primary antibodies were used for immunoblotting at 1:1000 dilution unless specified otherwise: anti-RIPK1 (3493), anti-phospho-RIPK1 (human) (65746), anti-phospho-RIPK1 (murine) (31122), anti-RIPK3 (human) (13526), anti-phospho-RIPK3 (human) (93654), anti-phospho-RIPK3 (murine) (57220), anti-IκBα (9242), anti-phospho-IκBα (2859), anti-NF-κB (6956), anti-phospho-NF-κB (3033), anti-MK2 (3042), anti-phospho-MK2 (3007), anti-p38 (9212), anti-phospho-p38 (9215), anti-JNK (9252), anti-phospho-JNK (4668), anti-caspase-8 (9746), anti-CYLD (8462) all from Cell Signaling Technology (Frankfurt, Germany), anti-MLKL (MABC604) from Merck Millipore GmbH, anti-SHARPIN (14626-1-AP) from Proteintech (Manchester, UK), anti-TRADD (610572) from BD Biosciences (Wokingham, UK), anti-human phospho-MLKL (ab187091), anti-murine phospho-MLKL (ab196436), anti-TNFR1 (ab205718) all from Abcam (Berlin, Germany), anti-HSP90 (ADI-SPA-835, dilution 1:3000) from Enzo Life Sciences, anti-RIPK3 (murine) (2283, dilution 1:3000) from ProSci (via Hölzel Diagnostika), anti-FADD (sc-6036), anti-caspase-8 (sc-6136) both from Santa Cruz Biotechnology (Heidelberg, Germany), anti-β-Actin (A5441), anti-FLAG (F3165) both from Sigma-Aldrich and anti-HA (11867423001) from Roche Pharma AG (Reinach, Switzerland).