ServicesFAQ
What happens when we send a sample to the core facility?
Your sample gets logged in our system and queued for production. The queue is a maximum of two weeks before the project start. We tend to start things in batch runs of 10 antigens at a time.
We run a single batch run every week. We run an SDS gel and transfer to nitrocellulose, stain it with Ponceau S and check that the protein is migrating where it is supposed to and that the concentration is sufficiently high for us to proceed. This blot will also be used for testing clones isolated by the first two screening protocols.
Next we immunize the animals with your target antigen. Currently we immunize using an adjuvant developed in-housewhich requires only 4 immunizations before fusion.
14 days after the first immunization each animal will be boosted.
Ten days after the boost we check the serum titre to see if each mouse has raised an immune response against your antigen.
Two weeks after the fist boost (the second immunization) the animals are boosted again and ten days later the serum is checked for titre.
Just before the fusion the mice will be immunized for the third time and the fusion carried out.
There is a 12 day incubation period while the cells grow and secrete antibody into the culture medium.
We change out the medium completely to remove antibodies secreted by unfused lymphocytes (which die naturally after about 5 days leaving only the hybridomas). This cuts down on background and false positives.
After about 12 days we consolidate the supernatants into 384-well plates and then use a single chip coated with your antigen as a kind of nano-ELISA.
Positives from this first round of screening are tested by conventional ELISA and by Western blot.
These screens take about a week to complete.
Assuming that we manage to isolate clones for you, we will then expand the cells and harvest about 10ml of usable culture supernatant for you to test and characterize. We will freeze down two aliquots of cells from each of the clones, one of which we will send to you and one we will bank in the core facility.
What if I need more of a particular antibody?
We offer a high volume ascites service which takes about six weeks and yields afrom 10-200 milligrams of antibody. Usually this does not need to be purified. Please check in the services link on the sidebar.