β actin catalog no sc47778

The reagents fenbufen, N,N’-dicyclohexylcarbodiimide (DCC), N-hydroxysuccinimide (NHS), and gadolinium (III) chloride hexahydrate (GdCl₃∙6H₂O) were purchased from Sigma Aldrich (St Louis, MO, USA). All other chemical reagents and solvents were obtained from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan) and Duksan Pure Chemical Co., Ltd. (Ansan, Republic of Korea). Tri-tert-butyl 2,2′,2″-(10-(2-((2-aminoethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (DO3A-^tBu-NH₂) was prepared according to the methods described in the literature [45 (link)]. λ-Carrageenan, PA, and other reagents were obtained from Sigma Aldrich (St Louis, MO, USA). Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Laboratories (Kumamoto, Japan). Antibodies for COX-2 (Catalog No. 12282) were purchased from Cell Signaling Technology (Beverly, MA, USA), and β-actin (Catalog No. sc47778) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

The cellular proteins were prepared from cells growing on a 6-well plate after treatments of BNAG1, or BNAG2 or NAG using a commercial RIPA buffer (5X) (Catalog No. AAJ62524AD, Thermo Fisher Scientific) containing a protease inhibitor cocktail (Catalog No. sc-29130, Santa Cruz Biotechnology Inc., Dallas, TX, USA) and 1 mM phenylmethylsulfonyl fluoride (Catalog No. sc-482875, Santa Cruz Biotechnology Inc.). Then the protein concentration was determined using the Bradford protein assay kit (Catalog No. 5000201, Bio-Rad, Hercules, CA, USA). SDS-polyacrylamide gel electrophoresis, protein transfer to nitrocellulose membranes (Catalog No. 88025, Thermo Fisher Scientific), incubation of membranes with primary antibodies against mouse iNOS and Cox-2 (Catalog No. NB300-605 and NB110-1948, both from Novus Biologicals, LLC), and horseradish peroxidase (HRP)-conjugated secondary antibody (Catalog No. 7074S, Cell Signaling), and visualization of protein bands were performed according to the procedures described previously [18 (link)]. β-actin (Catalog No. sc-47778, Santa Cruz Biotechnology Inc.) was used as the loading control. All Western blot assays were conducted in duplicates.

The cell culture medium RPMI 1640, DMEM, McCoy’s 5a, DMEM/F12, EGF, horse serum and fetal bovine serum were purchased from GIBCO (Invitrogen, Carlsbad, CA, USA). Antibodies against pAnxA2-Y23 (Catalog no. sc-135752), and β-actin (Catalog no. sc-47778) were obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA), whereas pAnxA2-S25 (Catalog no. PA5-37474) was from Thermo Fisher Scientific (Life Technologies Corporation, Grand Island, NY, USA). AnxA2 (clone 5; Catalog no. 610069) antibody was purchased from BD Pharmingen (BD Biosciences, San Jose, CA, USA). GFP antibodies for immunoblotting were purchased from Cell Signaling (Catalog no. 2955; Cell Signaling Technology, Inc., Danvers, MA, USA) and for immunoprecipitation experiments were purchased from Developmental Studies Hybridoma Bank, Iowa City, IA (Catalog no. DSHB-GFP-12A6). Protein A/G-agarose was acquired from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and immunoblot stripping buffer from Pierce Co. (Rockford, IL, USA). All other reagents and biochemicals were purchased from Sigma-Aldrich.