Il 6 enzyme linked immunosorbent assay elisa kits

Unless otherwise stated, all chemicals were obtained from the Sigma Chemical Company, St. Louis, MO, United States. Mouse anti-rat TfR1 (transferrin receptor 1) and fetal bovine serum (FBS) were purchased from Invitrogen, Carlsbad, CA, United States; rabbit polyclonal anti-mouse Fpn1 from Novus Biologicals, Littleton, CO, United States; rabbit polyclonal anti-Ft-L and anti-DMT1 (divalent metal transporter 1, SLC11A2) from Protein-tech, Chicago, IL, United States; and rabbit polyclonal anti-phospho-STAT3 (Tyr705) and mouse monoclonal anti-STAT3 both from Cell Signaling Technology, Inc., Danvers, MA, United States. Recombined mouse IL-6 protein was obtained from Gene Script, Piscataway, NJ, United States; IL-6 enzyme-linked immunosorbent assay (ELISA) kits from R&D Systems, Minneapolis, MN, United States; goat anti-rabbit or anti-mouse IRDye 800 CW secondary antibodies from LI-COR Bio Sciences, Lincoln, NE, United States; TRIzol reagent from Life Technologies, Carlsbad, CA, United States; AevertAid First Strand cDNA Synthesis Kit from Thermo Scientific, Waltham, MA, United States; FastStart Universal SYBR Green Master and LightCycler96 from Roche, Nutley, NJ, United States; and BCA protein Assay kit and protein RIPA lysis buffer from Beyotime Institute of Biotechnology, Haimen, JS, China.

Oyster β-thymosin was synthesized based on the previous study [18 (link)]. Human β-thymosin and LPS (Escherichia coli) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS) and Dulbecco’s modified eagle medium (DMEM) were purchased from Cellgro Mediatech, Inc. (Manassas, VA, USA). PGE₂, TNF-α, IL-1β and IL-6 enzyme-linked immunosorbent assay (ELISA) kits were purchased from R&D Systems Inc. (Minneapolis, MN, USA). Most of the antibodies were obtained from Cell Signaling Technology Inc. (Danvers, MA, USA): iNOS (D6B6S) Rabbit mAb #13120, COX-2 (D5H5) XP® Rabbit mAb #12282, TNF-α Antibody #3707, NF-κB pathway: NF-κB Pathway Sampler Kit #9936 (except p-NF-κB), β-Actin (D6A8) Rabbit mAb #8457, anti-rabbit IgG, horseradish peroxidase (HRP)-linked Antibody #7074, anti-mouse IgG, HRP-linked Antibody #7076, anti-rat IgG, HRP-linked Antibody #7077 (Beverly, MA, USA) and p-NF-κB(p-NFκB p65 (27.Ser 536): sc-136548), IL-1β(IL-1β (B122): sc-12742), and IL-6(IL-6 (10E5): sc-57315) antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Lipopolysaccharide (LPS) from Escherichia coli serotype (O111:B4) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum, streptomycin, and penicillin were obtained from Invitrogen Gibco (Grand Island, NY, USA). Griess reagent was purchased from Invitrogen, Thermo Fisher Scientific, Inc. (Eugene, Oregon, USA). TNF-α and IL-6 enzyme-linked immunosorbent assay (ELISA) kits were purchased from R&D Systems, Inc. (Minneapolis, MN, USA). Purchases of iNOS, COX-2, p-NF-κB p65, NF-κB p65, pI-κB, I-κB, p-ERK, ERK, p-JNK, JNK, p-p38, p38, and β-actin antibodies were made from Cell Signaling Technology (Beverly, MA, USA) and Thermo Fisher Scientific, Inc. (Rockford, IL, USA).