Specifications
Our Monoclonal Packages cover Three Phases:
Phase I (Immunization): Five BALB/c mice are immunized and their titers tested each week by ELISA. Immunization continues until titers are high enough for the project to move on to Phase II.
- boost with 25µg
- 1st boost @ day 0
- 2nd boost @ day 21
Phase II (Fusion with myeloma cells): The mouse spleen is removed and fused to mouse myeloma cells to create hybrids.
- ratio: 1 myeloma for every 4 B cells
- only hybridoma will grow, non-fused cells will die
Optional: Screening of positive wells allows the customer to determine which ELISA positive clone(s) they want used for expansion by sending the supernatant of positive wells for evaluation. (See below)
Phase III a (Hybridoma Subclone Stabilization): Positive wells are selected and subcloned several times to further isolate positive clones. We guarantee three finished hybridomas for the customer.
- positive wells selected, the 20 best wells are moved to 24 or 48 well plates
- ≥ wells from 24 plate is selected by ELISA & each is grown on a 96 well plate
- select the wells with a single cell
- do another subcloning with a single cell
- perform an ELISA on single cells & grow the cells up as a single clone
Phase III b (Optional Screening by WB or ICC): Positive wells are selected and screened by Western Blotting (WB) or Immunocytochemistry (ICC)
Below is the summary of the available Packages: -
| Package Name |
Specifications
|
| Peptide Synthesis & Conjugation |
Phase I: Immunization of 5 mice
(10-16 weeks) |
Phase II: Fusion of mouse spleen with myeloma cells
(2-3 weeks) |
Phase IIIa:
Screening of clones by ELISA & subcloning of positive wells
(4-6 weeks) |
Phase IIIb: Screening of positive wells by Western Blotting (WB) or Immunocytoche-mistry (ICC)
|
Final Clone Isotyping |
| MAb-MAS1
|
Immunogen provided by researcher |
√ |
√ |
3 positive clones
|
- |
√ |
| MAb-MAS1S |
√ (WB or ICC) |
| MAb-MAS2
|
Up to 25 aa, >90% HPLC
|
- |
|
MAb-MAS2S
|
√ (WB or ICC) |
|
