The mice were lavaged with 1 mL phosphate buffered saline (PBS), intraperitoneally injected, under anesthesia, and the bronchoalveolar lavage fluid (BALF) was obtained. After centrifugation at 300 × g for 10 min at 4°C, the supernatant was obtained and preserved at −80°C for TNF-α and total protein analyses. The total protein concentration in BALF was detected with a commercial bicinchoninic acid (BCA) assay kit following the manufacturer's instructions (Beyotime, Shanghai, China). The TNF-α levels were measured by an enzyme-linked immunosorbent assay (Nanjing KeyGen Biotech. Co., Ltd., Nanjing, China). The sediment was treated with erythrocyte lysate (Beyotime) for 10 min, and then centrifuged at 300 × g for 10 min at 4°C. Subsequently, the cells were resuspended in 500 μL PBS and observed using an inverted microscope (Leica DMi8, Leica Microsystems, Germany).
Enzyme linked immunosorbent assay
Manufactured by Keygen Biotech
Sourced in China
The enzyme-linked immunosorbent assay (ELISA) is a laboratory technique used to detect and quantify specific proteins or other molecules in a sample. It utilizes antigen-antibody interactions and an enzyme-based color change reaction to measure the presence and concentration of the target analyte.
Automatically generated - may contain errors
2 protocols using enzyme linked immunosorbent assay
1
Bronchoalveolar Lavage Fluid Analysis
2
Cytokine and Enzyme Measurement in Rat Serum
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The methods used to measure cytokine levels were described in our previous study [21 (link)]. Rat serum was centrifuged and prepared to evaluate interleukin (IL)-1β, IL-18, and tumor necrosis factor (TNF)-α levels by enzyme-linked immunosorbent assay according to the manufacturer's instructions (KeyGEN Biotech Company, Nanjing, China). The optical absorbance was detected by using a microplate reader (Bio-Rad, Hercules, CA, USA). Intestinal β-hexosaminidase level was measured using a colorimetric method according to our previously described method [18 (link)].
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