According to a previously described method (23 (link)), BALF was collected by flushing 1 ml ice-cold PBS back and forth three times through a tracheal cannula, and then centrifuged at 1,000 × g at 4°C for 10 min. The protein concentrations of IL-6 (cat. no. BMS603-2), IL-8 (cat. no. EMCXCL15), TNF-α (cat. no. BMS607-3), IL-17 (cat. no. BMS6001), IL-18 (cat. no. BMS618-3) and IL-1β (cat. no. EM2IL1B2) in the supernatant were measured using murine cytokine-specific ELISA kits (Invitrogen Life Technologies; Thermo Fisher Scientific, Inc.) in strict accordance with the manufacturer's protocol. The quantities of total protein (cat. no. ABIN996404), albumin (cat. no. ABIN2756308) and KGF (cat. no. ABIN2703018) in the BALF were measured as markers of epithelial permeability using ELISA kits (Antibodies-Online GmbH).
Protocol detail
BALF Cytokine and Protein Analysis
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Albumin
Antibodies
Cannula
Cold
Cytokine
Elisa
Il 17
Il 18
Il 1β
Il 6 protein
Murine
Permeability
Protein
Tnf α
Tracheal
Corresponding Organization :
Other organizations : Soochow University, First Affiliated Hospital of Soochow University
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Variable analysis
independent variables
- No independent variables explicitly mentioned.
- Protein concentrations of IL-6, IL-8, TNF-α, IL-17, IL-18, and IL-1β in the BALF supernatant
- Total protein, albumin, and KGF quantities in the BALF
- Collection of BALF by flushing 1 ml ice-cold PBS back and forth three times through a tracheal cannula
- Centrifugation of BALF at 1,000 × g at 4°C for 10 min
- Use of murine cytokine-specific ELISA kits (Invitrogen Life Technologies; Thermo Fisher Scientific, Inc.) to measure cytokine protein concentrations
- Use of ELISA kits (Antibodies-Online GmbH) to measure total protein, albumin, and KGF quantities in the BALF
- No positive controls mentioned.
- No negative controls mentioned.
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