Opteia mouse ige elisa set

Upon whole blood collection via cardiac puncture, the serum was isolated by centrifugation at 1,500 rpm for 20 min at 4°C. Determination of serum IgE levels was performed using the BD OptEIA™ mouse IgE ELISA set (#555248, BD Biosciences, CA, USA), according to the manufacturer's protocol. After the final step, plate was read with a microplate reader (VersaMax, Molecular Devices, CA, USA) at 450 nm absorbance.

At one day before the injection of OVA/Al(OH)₃, blood samples were collected from the tail vein. On the final day, whole blood was collected by cardiac puncture. The blood samples were incubated at room temperature for 1 h at 4 °C overnight and were centrifuged at 10,000 rpm for 10 min to obtain the serum. The amount of total IgE, OVA-specific IgE and IgG1 was measured in serum by using BD OptEIA Mouse IgE ELISA Set (BD Bioscience, San Jose, CA, USA), DS Mouse IgE ELISA (OVA) (DS Pharma Biomedical, Osaka, Japan), Anti-Ovalbumin IgG1 (mouse) ELISA kit (Cayman Chemical, Ann Arbor, MI, USA), respectively.

Serum concentrations of corticosterone were determined using the Corticosterone ELISA kit (Abnova). Samples were diluted at 1:50 and the assay was performed according to the manufacturer’s instructions. Serum concentrations of IGF-1 and IGFBP-3 were determined using the Mouse/Rat IGF-1 Quantikine ELISA kit (R&D Systems) and IGFBP-3 DuoSet ELISA kit (R&D Systems), respectively. Samples were diluted at 1:500 and the assay was performed according to the manufacturer’s instructions. Serum concentrations of total IgE were quantified with BD OptEIA™ Mouse IgE ELISA Set (BD Biosciences, San Chose, Canada) with samples diluted 1:100 according to the manufacturer’s instructions. Samples were assayed in monoplicate.

TNF-α, and total and specific IgE levels from lung homogenates or blood sera were measured by means of enzyme-linked immunosorbent assay (ELISA) with commercially available materials. Briefly, TNF-α and total IgE levels were assayed with Mouse TNF-α DuoSet (R&D Systems) and BD OptEIA Mouse IgE ELISA Set (BD Pharmingen, San Diego, CA), respectively. Specific IgE levels were measured by modified-sandwich ELISA [17 (link)]. Briefly, the plates were coated with a purified rat anti-mouse IgE, and sera were used as test samples. Biotinylated-OVA and peroxidase-conjugated streptavidin were reacted in order. The reactions were read at 450 nm.Leptin and adiponectin levels from lung homogenates or blood sera were measured by means of ELISA with commercially available materials, Mouse Leptin and adiponectin DuoSet (R&D Systems), respectively.