Mouse elisa kit

Serum levels of IL-6, TNF-α (EMC004.96 and EMC102a.96, Neo Bioscience Technology, Shenzhen, China) were measured by Mouse ELISA Kits according to the manufacturer’s protocols. Reagents, samples, and standards were prepared as instructed. After adding 50 μl standards and serum samples per well, the 96-well plate was incubated for 2 hours at 37°C. Then the liquid was removed and washed. And then 100 μl of Biotin antibody was added to each well before incubating for 1 h at 37°C, and followed by washing the plate 5 times with a washing buffer. 100 μl of HRP-avidin was added to each well, incubated for 1 hour at 37°C, and washed 5 times. After adding 90 μl TMB substrate to each well, the plate was protected from light and incubated for 20 minutes at 37°C. Color formation was stopped by 50 μl stop solution, and the optical density (OD) value was read at wavelength of 450 nm on a plate microplate reader within 5 minutes. Corresponding concentrations were converted from OD values according to the standard curve. Each serum sample was tested twice and the average value was taken for analysis.

Hematoporphyrin monomethyl ether (HMME) was obtained from Shanghai Yuanye Bio‐Technology Co., Ltd (China). Iron chlorides anhydrous (FeCl₃), methanol, triethylamine (TEA), N,N‐dimethylformamide (DMF), dihydroartemisinin (DHA), tannic acid (TA), methylene blue (MB), TMB were purchased from Aladdin Bio‐Chem Technology Co., Ltd (Shanghai, China). Cell counting kit‐8 (CCK‐8) was bought from APExBIO (USA). Simulated gastric fluid (SGF, USP) was purchased from Leagene Biotechnology (China). Brain‐heart infusion (BHI) and Columbia blood plates were obtained from Huan kai Guangzhou Microbial (China). Singlet oxygen sensor green (SOSG) and live/dead BacLight bacterial viability kit (L‐7012) were bought from Invitrogen (USA). BCA protein assay kit was purchased from Beyotime (China). Mouse ELISA kits were purchased from NeoBioscience Technology Co, Ltd (China). H. pylori strain ATCC 43504 and ATCC 700392 were obtained from American Type Culture Collection (ATCC, USA). LQ2# and CS01 were clinical isolates gifted by Prof. Meicun Yao (Sun Yat‐sen University, Shenzhen, China.). BALB/c female mice (6–8 weeks old) were purchased from Gempharmatech Co., Ltd (China). High‐purify water (Millipore Milli‐Q grade) with a resistivity of 18.2 MΩ was used in all the experiments. All reagents used in the experiments were analytical grade without further purification.

The levels of IL-1β, CXCL-1, IL-8, TNF-α, TGF-β, MMP-9, IL-10, and NE activity in BALF were measured using mouse ELISA kits (Neobioscience) according to the manufacturer’s instructions.

Based on the manufacturer’s specifications, the CXCL-1, IL-1β, IL-1RN, IL-8, IL-10, TNF-α, MMP-9, and TGF-β levels in BALF were detected with mouse ELISA kits (Neobioscience).

The animal blood sample was centrifuged at 3,000 rpm for 20 min to separate serum. According to the manufacturer's protocol, the levels of TNF-α, IL-1β, and IL-6 in serum were detected by the commercially available mouse ELISA kits purchased from Neobioscience (Shenzhen, China), and the serum of each animal was determined in duplicate.