Briefly, microtiter plates were coated for 24 h with the samples diluted 1 2 in PBS with 5% albumin.
In contrast to other studies, the materials were coated for the first time on 300 mm wafers and exposed on a 50 kV VISTEC SB3050DW variable shaped electron beam direct writer in a near-production environment at Fraunhofer CNT.
The plates were coated for 2 hours before cells were added.
In separate experiments, 24-well plates were coated for 2 h with 10 µg/mL human recombinant VCAM-1 or ICAM-1.
Briefly, flat bottomed 96-well polystyrene plates were coated for 18 h at 4°C with 21 nM human fibrinogen (Diagnostica Stago, Asnières, France) diluted in phosphate-buffered saline (PBS) (150 mM NaCl, 10 mM sodium phosphate, pH 7.2).
The primers were stained, the flow-through chambers were disassembled, and the BeadChips were coated for protection.
For competitive ELISA, Nunc maxisorb plates were coated for 2 h with DR4-Ig (100 ng per well) in 0.1 M sodium carbonate/bicarbonate buffer (pH 8.6) and the remaining binding places subsequently blocked with 2% BSA for 1 h.
For stimulation of effector and naïve T cells by anti-CD3/anti-CD28 antibodieSerotectec), 96-well plates were coated for 2 h at room temperature with 5 µg/ml of each antibody in phosphate buffered saline.
Ninety-six-well plates were coated for 18 h at 4 °C with 5 μg/mL KLH (Biosyn, Carlsbad, CA, USA) in PBS in a total volume of 120 μL per well, and wells were subsequently blocked with skimmed milk powder.
