Antibody testing - what if antigen is not available in a soluble form? (Sep/21/2005 )
Hi,
I want to generate antibodies against a protein that is difficult to solubilize. The QIAexpressionist manual says its OK to immunize mice with proteins in urea-containing buffer (IS THAT SO?). However, if that is indeed OK and I do start immunizing animals, how do I test the Ab titer of the serum?
Normally an ELISA is done with a range of antigen concentrations. If my antigen is urea-solubilised, I can't use it for ELISA right? That being the case, is there any other option whereby I can immobilise the antigen such that I can test my antibody?
I suppose one way is to get protein from mammalian cells and run it on a gel, do a western, but not only is this a long procedure, but also may not give a good idea regarding titre as such.
What are your opinions, and are there alternatives?
Thanks very much.
I want to generate antibodies against a protein that is difficult to solubilize. The QIAexpressionist manual says its OK to immunize mice with proteins in urea-containing buffer (IS THAT SO?). However, if that is indeed OK and I do start immunizing animals, how do I test the Ab titer of the serum?
Normally an ELISA is done with a range of antigen concentrations. If my antigen is urea-solubilised, I can't use it for ELISA right? That being the case, is there any other option whereby I can immobilise the antigen such that I can test my antibody?
I suppose one way is to get protein from mammalian cells and run it on a gel, do a western, but not only is this a long procedure, but also may not give a good idea regarding titre as such.
What are your opinions, and are there alternatives?
Thanks very much.
Anyone...?
pls...? I have heared once, that mice were immunized orally with inclusion bodies; But I do not know, is it a good idea, and what were the results of that experiments.