How much antigen is needed for immunizations? Approximately 1.2 mg in PBS
How much antigen is needed for Elisa screening? Approximately 3.0 mg in PBS
Can I immunize with peptides? Yes, however it might be helpful to couple the peptide to a carrier protein such as keyhole limpet hemocyanin, bovine serum albumin, or ovalbumin. It is also helpful to know if the protein has been immunogenic in rabbits, that is, if a polyclonal serum has been produced
Can I immunize with a protein coupled to a GST tag? Yes, but for Elisa screening, it is best to have the protein coupled to a different tag.
What volume is injected into each mouse? 200 ul/injection, 100ug of which is Ag
At what concentration should the antigen be? 1 mg/ml in saline or PBS is preferred
What is the Final Boost? Soluble Ag, injected i.v.
How many clones can be expected? From one to as many as 75+; this cannot be predicted
How long does a typical monoclonal project take? About 4-5 months
What is the source of antibody-secreting cells to be fused? The spleen
What is the fusion partner? Sp2/0, a myeloma line of Balb/c mouse origin, or mIL6SP2/0, which secretes the growth factor murine, mIL6