I'm interested in the insulin signaling pathway and want to identify markers of insulin resistance in DIO mice (e.g. pAKT/pIRS1/pJNK).

Normally, we perform euthanasia by pentobarbital overdose however I was informed that this drug, as well as several other anesthetic agents, can alter the phosphorylation state of proteins.

Can someone with experience tell me what is the optimal method of mouse euthanasia when studying changes in phosphorylation?

We are currently considering cervical dislocation. Someone had suggested decapitation following isoflurane anesthesia however several articles have reported that isoflurane can alter the phosphoproteome.

Thank you!