Anti igg1 antibody

Cancer cell lines (100,000 cells/tube) were incubated with antibodies against EpCAM (Miltenyi, Biotec, Custom antibody) and PD-L1 (XP version, Cell Signaling Danvers, MA) for 1 h on ice. Anti-IgG1 antibody was used as negative control (Abcam, Cambridge, UK). After 3 washes in PBS, samples were incubated with PE- or FITC-conjugated secondary antibodies for 1 h on ice. FACS analysis was performed by FACS-Canto instrument (BD, Becton-Dickinson, Franklin Lakes, NJ, USA).

ADP, collagen and thrombin were purchased from Probe&Go (Osburg, Germany). Bovine serum albumin (BSA, Fraction V) was purchased from GE Healthcare (Buckinghamshire, UK). The phospholipase C (PLC) inhibitor U-73122, protein kinase C (PKC) inhibitor chelerythrine, BAPTA-AM, fibrinogen, fibronectin, EGTA and cytochalasin B were purchased from Sigma (Vienna, Austria). Monoclonal anti-CD36 antibody [clone FA6-152] (ab17044), polyclonal anti-SR-A antibody (ab123946) and anti-IgG1 antibody were purchased from Abcam (Cambridge, UK), polyclonal anti-SRBI antibody (NB400-101) from Novus biological (Cambridge, UK).
The superoxide dismutase mimetic MnTMPyP pentachloride was from Santa Cruz (California, USA). Assay buffer used in P-selectin expression staining was Dulbecco’s modified phosphate-buffered saline (PBS; with or without 0.9 mM Ca²⁺ and 0.5 mM Mg²⁺; Invitrogen, Vienna, Austria). The antibodies against P-selectin, CD40L, TF and IgG1 were purchased from Becton Dickenson (Vienna, Austria). Fixative solution was prepared by mixing 30 ml FACS-Flow with 9 ml aqua dest. and 1 ml CellFix.

Influenza antigen-specific antibodies in mouse serum were titrated with enzyme-linked immunosorbent assay (ELISA). Briefly, the plates were coated with 4 μg/mL Fluzone^® 2017/2018 influenza vaccine (1 μg/mL each Influenza antigens), incubated with 10000x diluted mouse serum after blocking with 5% bovine serum albumin (BSA) in phosphate-buffered saline (PBS), washed, and subsequently incubated with horseradish peroxidase(HRP)-conjugated anti-IgG (1:10000; Abcam, Cambridge, MA, USA), anti-IgG1 antibody (1:10000; Abcam), or anti-IgG2a antibody (1:10000; Abcam). After sufficient color development of the substrate solution, absorbance was measured at 405 nm by a microplate reader.