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
MAS1

 

Immunogen provided by researcher 3 positive clones

 

-
MAS1S √ (WB or ICC)
MAS2

 

Up to 25 aa, >90% HPLC

 

-

MAS2S

√ (WB or ICC)