Bovine serum albumin (bsa)

DHEA (Form I) [28 (link)] (purity > 99%) was purchased from Shanghai Macklin Biochemical Co., Ltd., Shanghai, China. while coformers (purity > 98%) and polymers (purity > 98%) were both obtained from Shanghai Aladdin Biochemical Technology Co., Ltd., Shanghai, China. All solvents at analytical grade and bovine serum albumin (BSA) (purity > 98%) were bought commercially from Sinopharm Chemical Reagent Co., Ltd., Shanghai, China. All chemicals were used without further purification in this study. Besides, the pH 2.0 buffer in this study consisted of 0.2 mol/L hydrochloric acid and 0.2 mol/L phosphate. In contrast, in the pH 4.5 solution, hydrochloric acid was replaced by citric acid monohydrate to adjust the pH value of this system. As for the pH 6.8 buffer, it was made up of 0.2 mol/L phosphate solution.

Bovine serum albumin (BSA) was purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Copper nitrate and sodium hydroxide were purchased from Aladdin Reagent Co., Ltd. (Shanghai, China). Na₂S (anhydrous) was purchased from J&K Scientific (Shanghai, China). Copper standard solution was obtained from Beijing North Weiye Metrology Technology Institute (Beijing, China). An enhanced BCA protein assay kit was purchased from Beyotime (Shanghai, China).

Bovine serum albumin (BSA) and sodium hydroxide (NaOH) were purchased from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). Iridium trichloride (IrCl₃) was acquired from Aladdin Reagent Co. (Shanghai, China). Doxorubicin hydrochloride (DOX, purity 98%), phosphate buffered saline (PBS), fetal bovine serum (FBS), calcein acetoxymethyl ester (Calcein AM) and propidium iodide (PI) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Cell counting lit-8 (CCK-8) was ordered from Dojindo Laboratory (Dojindo Co., Ltd. Japan). Dulbecco’s modified Eagle medium (DMEM), penicillin, and streptomycin were obtained from Gibco (Invitrogen, Carlsbad, CA). The Human osteosarcoma cancer cell line (Saos-2 cells) and A549 cells were obtained from the Institute of Basic Medical Sciences Chinese Academy of Medical Sciences (Shanghai, China). Milli-Q water was prepared using a Milli-Q system (Bedford, MA, America). All chemicals were used as received without further purification.

Mature lotus seedpods of Nelumbo nucifera Gaertn. (cultivar: Number 2 Wuhan plant) were obtained from Honghu Lantian (Hubei, China) in late July, 2012, and identified by Professor Xueming Ni from the Department of Botany, Wuhan Plant Institute of the Chinese Academy of Science. (+)-Catechin, ferulic acid, caffeic acid, syringic acid, m-coumaric acid, 3-hydroxybenzoic acid, 3-hydroxyphenylacetic acid and 3-hydroxyphenylpropionic acid were purchased from Sigma Chemical Co. (St. Louis, MO, USA). The β-glucuronidase demonstrated both β-glucuronidase (600 U/mg solid) and sulfatase (50 U/mg solid) activity. Methanol and formic acid (HPLC grade) were purchased from Fisher Scientific (Massachusetts, USA). Phosphate buffer saline (PBS, pH 7.4), bovine serum albumin (BSA) and d-glucose were purchased from Sinopharm Chemical Reagent (Shanghai, China). All other chemicals were of analytical grade.

The particle size distribution (PSD) of the bulk sludge was detected by a laser granularity distribution analyzer (S3500, Microtrac Instruments, Montgomeryville, PA, USA). The extracellular polymeric substances’ (EPSs) contents of the bulk sludge samples were regularly measured during the experiment. The heat treatment method was used for the EPS extraction [34 (link)]. The concentrations of COD and TSS were measured using the standard method [35 ]. PS was detected with the phenol-sulfuric acid method at 490 nm with glucose (GR, Sinopharm Group, Shanghai, China) as a standard and PN was analyzed using the Lowry–Folin method with Bovine Serum Albumin (BSA) (GR, Sinopharm Group, China) for the calibration at 750 nm [36 ,37 (link)]. An analyzer (BIOGAS 5000, Geotechnical Instruments Ltd., London, UK) and a wet gas flowmeter were used to monitor the composition and volume of the biogas. The dissolved organic matter (DOM) in influent, effluent, supernatant, and cake layer samples were further characterized by fluorescence spectra analysis (F-7000FL, HITACHI, Japan) [38 (link)].

Six porcine longissimus dorsi muscles (24 h post-mortem) and pork fat were randomly sampled at Sushi Meat Co., Ltd. (Huai’an, China), then frozen at −20 °C and were used within one week. Bovine serum albumin (BSA) and other chemicals were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China), and all chemicals were analytical grade.

Sodium succinate, rotenone, cytochrome c, coenzyme Q₁, coenzyme Q₁₀, antimycin A, decylubiquinol, 2′,7′-dichlorofluorescin diacetate (DCFH-DA) and Rhodamine 123 were purchased from Sigma (St. Louis, MO, USA). Tris base and NADH were purchased from Amersco Inc. (Palm Harbor, FL, USA). 2,6-Dichlorophenolindophenol (DCPIP) was purchased from Merck & Co. Inc. (Kenilworth, NJ, USA). Bovine serum albumin (BSA) for protein quantification was purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). All the other reagents used were of reagent grade and prepared in double-distilled water.