Anti-idiotype Monoclonal Antibody Protocol
|10 Balb/c mice or 6 rats or 2 rabbits|
|Complete Freund's Adjuvant (CFA) initially, followed by incomplete Freund's Adjuvant (IFA) or our proprietary immunization adjuvant for all subsequent injections except final boost.|
|5-10 mg purified antibody from customer. Whole molecule of the antibody will be used for immunization.|
|The immunogen is diluted with sterile saline and combined with the appropriate adjuvant. The antigen and adjuvant are mixed thoroughly to form a stable emulsion, which is injected intraperitoneally, providing enhanced immune response from the sustained presence of the immunogen.|
|Fusion Test bleed antisera samples are tested by ELISA with the target antibody and a negative control antibody with the same isotype (Provided by customer). Cell fusions are performed on the mice showing desired immune response to the immunogen.|
Expansion and Screening
|The fused cells are plated into 96-well culture plates and screened by ELISA with the target antibody and a non-related negative antibody for the specific clones. The desired clones are then expanded into 24-well plates and isotyped.|
Subcloning, expansion and cryopreservation
|The selected clones are then subcloned by limiting dilution until they are monoclonal and stable hybridomas. Two subclones from each parental clone are expanded into culture flasks, and 4-6 vials of cells for each subclonal cell line are cryopreserved.|
|For rabbit anti-idiotype antibody services, the selected clones are performed V-region sequencing.|
|The typical delivery includes two vials of frozen cells or one flask of live cells and 5 ml of culture supernatant per clone.|
We provide free cell line storage service for 6 months from the date of cell freezing. If customers require continued storage service after the end of six-month period, the charges are set at $38 per cell line per year.
Our customer service representatives are available 24 hours a day, Monday through Friday to assist you.