HI,

We have tried invivo stimulation phospho zap70 in mouse with the intraperitoneal administration of mouse CD3 antibody using 700ng and 10 ug. 10 and 30 after CD3 administration, we collect the whole spleeen and homogenization and LYsis was performed according PathScan® Phospho-Zap-70 (Tyr319) Sandwich ELISA Kit #7171 kit from Cell signaling technology. But we neither observed the activation over the basal nor over the blank.

Kindly suggest the protocol or activation and estimation of phospho zap70 in mouse spleen or whole blood by ELISA?

your inputs are valuable to set up the asasy.