If I were you, I would just make a gfp-tagged construct--this is an easy way to make sure that the protein is there, and made it to the membrane. If necessary, you can blot for gfp, or add other tags in the process for alternative blotting (one on each side to make sure it is not cleaved, for example.)
Jacob ----- Original Message ----- From: Qing Lu To: [email protected] Sent: Thursday, September 02, 2010 7:42 PM Subject: [ccp4bb] Confirming expression of a GPCR in HEK293 Dear All, Apologies for a non-ccp4 question. I am planning to express a GPCR in HEK293T cell line by transfecting with the requisite cDNA for performing an assay. I would like to confirm the expression of the protein in the cell line by western blot or histochemical studies. Any pointers in this regard would be helpful. Thanks, Qing Lu ******************************************* Jacob Pearson Keller Northwestern University Medical Scientist Training Program Dallos Laboratory F. Searle 1-240 2240 Campus Drive Evanston IL 60208 lab: 847.491.2438 cel: 773.608.9185 email: [email protected] *******************************************
