- Coating : 250 ng/well overnight a 4°c - washing : PBS tween 0,05 % - blocking : gelatin 0,5 % PBS Tween 0,05%, 1 heure à TA - washing : PBS tween 0,05 % - primary antibody dilution : 1/50 then following dilutions by 1/3 - incubation : 1 hour at room temperature - detection: anti-IgY-HRP, 1 hour at RT - substrate: TMB, 10 min at RT in the dark - Stop by : HCl 1N - OD reading at 450 nm