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2 protocols using anti il18 a1115

1

AMPK Signaling Pathway Modulation

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LPS O111:B4 (L2630), Doxycycline (D3447), 2-Deoxy-D-glucose (D8375) and compound C (P5499) were obtained from Sigma-Aldrich. Subcellular protein fractionation kit (78840) was obtained from Thermo Fisher. Metformin (S1950) and AICAR (S1802) were obtained from Selleck Chemicals. ATP (tlrl-atpl) was obtained from InvivoGen. StrataClean resin (400714) and chemical competent cells (200315) were obtained from Agilent.
For immunoblot, AMPKα1(#2795), AMPKα2 (#2757), AMPKα1/2 (#5831) and pT172-AMPK (#2535) were purchased from Cell Signaling Technology. Anti-Flag (F1804), anti-Flag (F7425) and anti-HA (H6908) were obtained from Sigma-Aldrich. Anti-GSDMD (ab219800) and anti-Caspase-1 (ab108362) were obtained from Abcam. Anti-β-Actin (sc-47778), anti-GST (sc-138) and anti-Na/K-ATPase α1 (sc-21712) were purchased from Santa Cruz Biotechnology. Anti-IL18 (A1115) was obtained from Abclonal Technology. phospho-GSDMD S46 was generated by Abclonal Technology.
For FCAS analysis, CD45-APC-Cy7(clone 30-F11), CD3-FITC (clone 145-2C11), CD8-PerCP-Cy5.5 (clone 53-6.7) and CD4-APC (clone RM4-5) were obtained from BioLegend.
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2

Complement Activation Pathway Profiling

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Bovine serum albumin (BSA) and Rabbit anti-BSA IgG were purchased from sigma-Aldrich Chemical Co (StLouis, MO, USA). Anti-C3a(JF10-30) and anti-C3aR (NBP2-1569) were purchased from Novus Biologicals (Littleton, CO, United States). Anti-C5a (ab194637), anti-C5aR (ab59390) and IL-1β (ab254360) were obtained from Abcam (Cambridge, MA, United States). Anti-p-JAK2 (3771s), anti-JAK2 (3230s), anti-p-STAT3 (9145s), anti-STAT3 (12640s), anti-SOCS3 (9021s), anti-p-IKKα/β (2694s), anti- IKKα/β (8943s), anti-p- NF-κB p65 (3033s), anti- NF-κB p65 (8242s) were obtained from Cell Signaling Technologies (Beverly, MA, USA). Anti-IL-18 (A1115) was purchased from ABclonal Technology (Wuhan, China). The complement C5a(HY-P7695) and STAT3 activator Colivelin TFA(CAF, HY-P1061A) were obtained from MCE (Shanghai, China).
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