Ab100705

ELISA kits for detection of IFN-α (42120-1), IFN-β (MIFNB0), IFN-γ (MIF00), TNF-α (MTA00B), IL-2 (M2000), IL-6 (M6000B), IL-10 (M1000B), IL-12 (M1270), and IL-23 (M2300) were purchased from R&D System. ELISA kits for detection of TGF-β (ab119557), IL-1α (ab199076), IL-1β (ab100705), and IL-4 (ab100710) were purchased from Abcam. Cytokine levels were determined using corresponding ELISA kits according to the manufacturer’s instructions.

On the 105th day of UVB irradiation, dorsal back skin tissue samples were obtained from the region around the gluteal area. Skin tissue homogenates were subjected to the measurement of IL-1β and IL-10 contents using mouse IL-1β (ab100705; Abcam, Cambridge, UK) and IL-10 (ab108870; Abcam, Cambridge, UK) enzyme-linked immunosorbent assay (ELISA) kits, following the manufacturer’s instructions. Optical density readings were taken at 450 nm using a microplate reader.

Enzyme-linked immunosorbent assay (ELISA) was used to quantify the hippocampal levels of IL-1β and TNF-α. In the in vivo experiments, isoflurane-anesthetized mice were decapitated 3 h after the LPS injection. Within 1 min after the decapitation, the hippocampi were extracted, quickly frozen, and stored at −70 °C until use. In the in vitro experiments, the slices collected after the LPS or saline exposure were used. Mouse IL-1β ELISA (ab100705, Abcam) and TNF-α ELISA (ab100747, Abcam) kits were used according to the manufacturer’s recommendations. Neural tissue was homogenized on ice in the extraction buffer recommended by the manufacturer (100 mM Tris, pH 7.4, 150 mM NaCl, 1 mM EGTA, 1 mM EDTA, 1 % Triton X-100, 0.5 % sodium deoxycholate) with 1 mg/mL of protease inhibitor cocktail (cOmplete, Sigma-Aldrich) and 0.01 mg/mL of phosphatase inhibitor cocktail (P5726, Sigma-Aldrich). The protein concentrations were determined using a BCA protein assay kit (Pierce, Rockford, IL). The absorbance at 450 nm was measured with an iMark Microplate Absorbance Reader (Bio-Rad).