Pierce bicinchoninic acid bca protein kit

Generally, all chemicals were obtained from FUJIFILM-Wako (Tokyo, Japan), unless stated otherwise. Fetal bovine serum (FBS) for cell culture was purchased from Gibco (GIBCO BRL, Grand Island, NY, USA). Cell Counting Kit-8 (CCK-8) and Cytotoxicity lactate dehydrogenase (LDH) Kit-WST were purchased from Dojindo Molecular Technologies (Rockville, MD, USA). Cellular ROS Kit (#ab113851) was obtained from Abcam (Tokyo, Japan). Nuclear Extraction (NE)-Kit and human TNF-alpha ELISA Ready-SET Go Kit (#88-7324) were obtained from eBiosciences (San Diego, CA, USA), respectively. Pierce bicinchoninic acid-(BCA) protein Kit and Enhanced chemiluminescent-(ECL) Western blotting substrate were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Bovine serum albumin-(BSA) fraction V was purchased from Roche (Mannheim, Germany). All antibodies for Western blot analysis were purchased from Cell Signaling Technology (Tokyo, Japan), except anti-Nrf2 (Proteintech Corp., Tokyo, Japan), anti-p65 NF-ĸB, and horseradish peroxidase (HRP)-conjugated second antibody (Sigma-Aldrich, Burlington, MA, USA), anti-rabbit (IgG) Alexa Fluor^TM 488-conjugate (Alexa-488), anti-rabbit (IgG) Alexa Fluor^TM 594-conjugate (Alexa-594) (Molecular Probes, Invitrogen, CA, USA). DAPI (4′,6-diamino-2-phenylindole) dye used for the nuclei staining was purchased from Molecular Probes (Eugene, OR, USA).

Total IgG was purified from SD rat serum using agarose protein G (Beyotime, Jiangsu, China). Pierce bicinchoninic acid(BCA) protein kit was used to detect IgG concentration (Thermo Fisher Scientific Inc, Waltham, US). The following chemicals were also used: DEN (Item No. N-0756, purchased from Sigma). Anti-p38 mitogen-activated protein kinase (p38 MAPK) antibody (8690T, CST); p-p38MAPK (4511T, CST); Bcl-2 (AF6139, Affinity); Bax (A00183, Affinity); β-actin (BM0627, BOSTER); and hematoxylin-eosin (HE, G1120, Sole Bauer).