Making antibodies - Anybody has designed a custom antibody? (Dec/05/2007 )
I'm working on a protein which doesn't have good commercial antibodies for IP. Therefore, I wanted to order a custom made antibody. Does any of you know, by any chance, if there are traps or details I should be wary of when choosing an epitope? Some commercial antibodies, which has been made on artificial peptides are not suitable for IP and ICH, and I'd like to have a better one.
Have a look at your protein and pick a region that's in a fairly open configuration - therefore more likely to bind.
I sent my protein sequence to the company and tell them if I want Ab specific for that sequence (species specific) or maybe for other species also. They suggested the possible peptides as antigens and I chose what I wanted.
The main problem to use peptide specific antibodies in IP is their antigen affinity. You'll need high affinity to capture your protein in IP. For your purpose, is better to immunize with pure protein. If not, you can synthetize any immunogenic peptide, immunize, purify polyclonal sera against peptide immobilized onto Sulfolink column, and use in IP.
I don't have the animals for immunization (and I am not sure if anyone could teach me in the institute, and I have heard it's very easy to make a mistake here), so I guess I'll just examine the epitopes, or synthesise my protein on e.coli (that would be long too...) and then pay some company to make antibodies for me (if I have budget! )