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 antibody development by immunization via the footpad will take 33 days and 55 days for immunization via subcutaneous injection. On day 33 or 55 and after our first screening by ELISA we can provide the user with a small amount of supernatant (50 ul maximum) for their own screening.
2) How much antigen should I provide?
For footpad immunization, 600 ug of antigen is required, ideally at a concentration of 1 mg/ml. 0.5 mg/ml is required for subcutaneous immunization. This amount includes the needs for screening.
3) What determines the type of immunization that will be done?
Mice will be immunized via the footpad if the immunogen is a primary or transfected cell line; purified protein can be used for subcutaneous immunizations.
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 screening and there is an additional cost (contact us for more information).
6) Do you perform western blott for screening?
No, but we can provide advise on how to proceed for screening with more than 100 samples and as few as 50 ul if western blotts are your goal.
7) Do you generate rat antibodies?
8) Do you generate rabbit antibodies?
9) How many clones will I receive?
You will receive all of the clones that show strong binding by ELISA, which can be up to 150. If it is needed, we can collect up to 2,000 clones (i.e. for marker discovery). Request a special quote for this service.
10) I don't have a purified protein or peptide for immunization; can I provide cells expressing the recombinant protein?
Yes. We can provide advice on protocols to generate those cells or put you in contact with Dr. Yutong Sun, Director of Functional Genomics Core (YSun2@mdanderson.org)
11) Which are the best 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?
Yes. Long Vien can provide stock from those cells.
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 do we need to provide?
The shortest peptide should be of 20 amino acids in length and conjugated to KLH, an antigenic molecule that helps to trigger the immune response. Details are provided during the initial discussion about your project.
Full length proteins (or longer peptides) do not need to be conjugated.