全血实验实验方案:1) 取肝素抗凝血用IMDM培养基1:10稀释;2) 加180ul稀释后血样到96孔板;3) 加入20ulLPS, 浓度梯度: 0.1, 0.3, 1, 3, 10 ng/mL; 时间点: LPS 刺激3小时, 6小时和过夜。4) 离心后取上清检测TNF-a等细胞因子.5) 细胞因子检测参照ELISA试剂盒操作手册。
Brief protocol of Whole blood assay:1) Blood sampled on heparin2) Dilute blood at 1/10 in IMDM media + Penicillin/Streptomycin 3) Add 180µl per well in 96 well plates.4) Add 20µl of LPS preparation in each well.5) Perform a LPS dose effect: 0.1, 0.3, 1, 3, 10 ng/ml6) Perform a kinetic of stimulation: 3H, 6H & ON.7) Centrifuge the supernatants and freeze at -80°C 8) Assess TNF release by ELISA