Mouse il 17a elisa kit

Blood was collected from NC/Nga mice on day 18, and serum samples were obtained by centrifugation (4,300 g for 5 min.). The total immunoglobulin E (IgE) concentration in serum was determined using the mouse IgE EIA kit (Yamasa Corp., Chiba, Japan), according to the manufacturer's instructions. Axial lymph nodes (LN) of each group of NC/Nga mice were removed, and lymphocytes from the LN were prepared 18 days after the first HLA-G1 treatment and stimulated with 1 µg/ml of immobilized anti-CD3 antibody (clone 145-2C11) and 5 µg/ml of soluble anti-CD28 antibody (clone 37.51) (BD Biosciences, San Jose, CA, USA) for 72 h. Supernatants were collected, and interferon (IFN)-γ, interleukin (IL)-13, and IL-17A concentrations were measured using the mouse IFN-γ Quantikine ELISA Kit (R&D Systems, Inc., Minneapolis, MN, USA), mouse IL-13 DuoSet ELISA Kit (R&D Systems, Inc., Minneapolis, MN, USA), and mouse IL-17A ELISA Kit (RayBiotech, Inc., Norcross, GA, USA), respectively, according to the manufacturer's instructions.