1) How long does it take to develop an antibody?
We perform two types of immunizations: a) foot pad injections and/or b) subcutaneous injection.
The total procedure for footpad will take 33 days and for subcutaneous 55 days. On day 33 or 55 and after our first screening by ELISA we can provide the user with small amount of supernatants (50 ul maximum) for their own screening.
2) How much antigen do I have to provide?
For footpad immunization, 500 ug of antigen is required, at the concentration 0.5 mg/mL and at least 0.1 mg/mL for subcutaneous immunization. This amount includes the needs for screening.
3) What determines the type of immunization that will be done?
The mice will be immunized at the footpad if the primary or transfected cell lines are the immunogen; the purified protein can be used for subcutaneous immunization.
4) What type of screening does the Core perform?
Primary screening will be done by ELISA.
5) Do you perform FACS?
Yes, only for secondary screenings and at an additional cost.
6) Do you perform WB for screening?
No, but we can provide you advise on how to proceed for screening with more than 100 samples and as few as 50 ul if WB is your goal.
7) Do you generate rat antibodies?
8) Do you generate rabbit antibodies?
9) How many clones may I receive?
All the clones that show strong binding by ELISA, which can be up to 100.
10) I don't have a purified protein or peptide for immunization; can I provide cells expressing the recombinant protein?
11) Which are the better cells to express my protein?
- Mouse fibroblast L cells
- CT39 T cell line
12) I don't have the mouse cells that you suggest to express my protein; can you provide me with them?
13) Does the Core test mycoplasma in the cells we provide?
If you cannot provide recent proof of mycoplasma screening, we can do it for you at an additional cost.
14) What minimal length of peptide we need to provide?
If the peptide is less than 30 amino acids, it needs to be conjugated with KLH (or other carrier protein).