Please help - FACS to ELISA for antibody testing
Posted 01 March 2012 - 04:11 PM
I did ELISA many times before but for secreted proteins - I mean cell based (like the hybridomas) and even estimated the expression titer. But now I have the cells which express proteins ON the surface and I need to assay with a commercially available antibody using ELISA.
To start with - how do I coat the plates?
Thanks for your replies.
ps: I also posted this same post in the ELISA group of this forum but no answer.
Posted 06 March 2012 - 02:36 PM
I am trying to find out how to assay my protein of interest using ELISA when the protein is ON the surface of the cell. Through online research, I understood that we need to lyse the cells. Looking for more info.
Posted 07 March 2012 - 03:11 AM
Alternatively, you could vigorously wash your cells, dialyze, concentrate the wash and test and/or lyse, dialyze, concentrate and test.
A question before starting is what concentration mg/ml, ug/ml, ng/ml, pg/ml are you expecting? Can you obtain some of the protein to test your elisa? You will need some sort of positive/negative control to determine if your test works.