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Table 1 Summary of assay conditions that were tested and selected for the autologous tumor IFN-γ ELISPOT assay

From: A modified human ELISPOT assay to detect specific responses to primary tumor cell targets

Item optimized Comparison Selected for assay
Plates Clear vs. Opaque Opaque
Targets (tumor cells) sCD40Lt stimulated vs. non-stimulated sCD40Lt stimulated
Effectors (PBMC) Overnight cultured vs 2 h cultured Overnight cultured cells
Effector:Tumor Ratio 100:100, 100:200, 100:300K 100:200K cells/well
  200:100, 200:200, 200:300K  
Incubation time of assay 24 and 48 hours 48 hour
Media 5% and 10% HuAB vs. 5% and 10% FBS 10% HuAB
Substrate Detection System AEC, TrueBlue™, and BCIP TrueBlue™