Mouse tnf α and il 6 elisa kits

Serum inflammatory cytokines were quantified by the Luminex 200 instrument (Merck Millipore, Billerica, MA, USA) using a Th1/Th2/Th9/Th17/Th22/Treg Cytokine 17-Plex Mouse ProcartaPlex™ Panel (Thermo Fisher Scientific). Cytokines in RAW264.7 cell supernatants were analyzed by mouse TNF-α and IL-6 ELISA kits (BD Biosciences, San Diego, CA, USA) as described previously [16] (link). For cAMP detection, cells and psoriatic skin biopsies were lysed and isolated by 0.1 M HCl and the intracellular cAMP level was quantified by using the direct cAMP ELISA kits (Enzo Life Sciences, The Netherlands) according to the directions.

We synthesized 3′,4′-dihydroxyflavone and flavone (Fig. 1A) using our previously reported procedure (An et al., 2017 (link)). LPS from Escherichia coli serotype O55:B5 (L6529) and luteolin (L9283) were purchased from Sigma-Aldrich (MO, USA). HyClone (GE Healthcare) culture media and other materials were used for cell culture. Rabbit antibodies against p38 (#9212S), phosphor-p38 (#9215S), JNK (#9252S), phosphor-JNK (#9251S), ERK (#9101L), phosphor-ERK (#9102), iNOS (#2982S) and COX-2 (#4842S) were purchased from Cell Signaling Technology (MA, USA). Antibodies against β-actin (SC-47778 HRP) and NF-κB (rabbit, SC-372) were purchased from SantaCruz Biotechnology (CA, USA). A rabbit antibody against anti-ionized calcium-binding adapter molecule 1 (Iba-1, #019-19741) was purchased from Wako (Tokyo, Japan). Secondary antibodies were purchased from Bio-Rad Laboratories (CA, USA), SantaCruz Biotechnology and Invitrogen (CA, USA). Mouse TNF-α and IL-6 ELISA kits were purchased from BD. A mouse IL-1β kit was purchased from Invitrogen.