Anti mouse cd16 cd32 antibody 2.4g2

Mice were immunized three times each with 10, 000 sporozoites administered by IV or ID in the presence or absence of laser illumination as detailed above. The mice were sacrificed 7 days after the final immunization and the liver, spleen, and blood were collected. The liver and spleen were dissociated by a 70 μm cell strainer. The cell suspensions, along with blood samples, were treated with Ammonium-Chloride-Potassium lysing buffer to remove red blood cells, and lymphocytes were isolated using Percoll (33%) as described [20 (link)]. Lymphocytes were then stimulated with 1mg / ml PyCSP280-288 peptide (SYVPSAEQI) for 21 hours at 37 °C with 1 μg / ml Golgin-plug in the culture for the final 5 hr. The stimulated cells were harvested, fixed with 2% formaldehyde, permeabilized with permeabilization buffer (eBioscience), and stained with indicated antibodies. Among the antibodies used, PerCP-Cy5.5 anti-Mouse CD8α antibody (clone 53–6.7) was purchased from eBioscience, PE anti-mouse CD11a Antibody (M17/4), Alexa 647 anti-mouse CD90.2 (30-H12) antibody, and FITC anti-mouse IFN-γ antibody (XMG1.2) from Biolegend, anti-mouse CD16/CD32 antibody (2.4G2) from BD Biosciences. The stained cells were assessed on FACSAria (BD Biosciences) and analyzed by FlowJo software (version 7.6.5).