Anti mhc 1 fitc

To investigate whether DENV-2 induces changes in molecules presented in the membrane of platelets, cytometric assays were carried out for CD62P, CD42b, MHC-I, and CD40L surface molecules. After treatment, the platelets were stained with specific antibodies for 30 min at RT in the dark and cells fixed with 1% paraformaldehyde solution. Antibodies used for these assays were conjugated to fluorophores as follows: anti CD62P-PE (eBiosciences, San Diego, CA, USA), anti CD42b-PE (BioLegend, San Diego, CA, USA), anti MHC-I-FITC (BD Biosciences, San Jose, CA, USA), and anti CD40L-PE (BD Biosciences, San Jose, CA, USA). For each treatment, 1 μg of fluorescent antibody was added to each 10⁶ platelets for incubation. Then platelets were washed and re-suspended in 250 μL of PBS for acquisition. The events corresponding in size (FS) and granularity (SS) to platelets were selected. Readings were made with a FACS ARIA-III (BD, San Jose, CA, USA) cytometer. A total of 150,000 events were acquired and platelets were selected as singlets. The assays were independently conducted in triplicate. Finally, the acquisition analysis was done on FlowJo 8.7 software (BD, Franklin Lakes, NJ, USA).

DEAE-52 cellulose was obtained from GE Healthcare (Uppsala, Sweden). The monosaccharides, comprising ribose (Rib), glucuronic acid (GlcA), glucose (Glc), galactose (Gla), mannose (Man), arabinose (Ara), rhamnose (Rha) and xylose (Xyl), lipopolysaccharide (LPS), active carbon, trifluoroacetic acids (TFA), and FITC-Dextran, were purchased from Sigma-Aldrich (St Louis, MO, USA). T-series dextrans (T-10, T-40, T-70, T-500 and Blue dextran) were purchased from Solarbio (Beijing, China). Fetal bovine serum (FBS) and Penicillin-streptomycin were purchased from MRC (Changzhou, China). Medium RPMI-1640 and phosphate-buffered solution (PBS) were purchased from Gibco (Grand Island, NY, USA). Granulocyte-macrophage colony-stimulating factor (GM-CSF) was purchased from PeproTech (Rocky Hill, NJ, USA). The other chemical reagents were purchased from Tianjin Fuchen (Tianjin, China).
The antibodies for flow cytometry comprising anti-CD40-APC, anti-CD86-APC, anti-CD11c-FITC, and ELISA kits including tumor necrosis factor-α (TNF-α), IL-1β, and IL-12p40 were purchased from Elabscience (Wuhan, China). Anti-MHC-I-FITC, anti-MHC-II-PE, and anti-CD11c-PE were bought from BD Biosciences (San Diego, CA, USA).