We have created custom mouse monoclonal antibodies for numerous applications including functional assays, immunoprecipitation, IHC, IF, flow cytometry, Western blot, and ELISA.
Peptide synthesis of up to 18 AA residues (20-30 mg) with a purity of >80% peptide conjugation to KLH
Recombinant protein preparation
Note: If there are no specific requests, 0.02% sodium merthiolate will be added to the antisera and antibody products.
ABclonal provides rapid immunizations within 2-5 weeks. This is suitable for the preparation of mouse monoclonal antibodies and polyclonal antibodies. Compared with conventional adjuvants, we have found our protocols to have a faster turnaround and to use less antigens.
We also provide comprehensive services for antibody cloning, engineering, expression, and purification in addition to custom antibody development.
ABclonal's standard option for mouse monoclonal antibody is optimized to produce antiserum in the shortest possible time. It is the most affordable option, popular among researchers.
The protocol uses five mice that are Specific-Pathogen-Free. Injections are subcutaneous (SQ) as emulsions in Complete Freund’s Adjuvant (CFA) or Incomplete Freund’s Adjuvant (IFA). We guarantee that the same antigen will be injected into all five animals.
|Control Serum Collection||Day 0||Pre-immune bleed (50-100 µl per mouse)|
|Primary Injection||Day 1||Immunize with 0.50 mg of antigen in CFA, 10 SQ sites|
|1st Booster||Day 14||Boost with 0.25 mg of antigen in IFA, 4 SQ sites|
|2nd Booster||Day 28||Boost with 0.25 mg of antigen in IFA, 4 SQ sites|
|Serum Collection||Day 35||Test-bleed (50~100 µl per mouse)|
|3rd Booster||Day 42||Boost with 0.25 mg of antigen in IFA, 4 SQ sites|
|Serum Collection||Day 56||Send bleeds to customer for validation (50~100 µl per mouse)|
|ELISA Test||Day 60||
ELISA titration (results available online);
Verify disposition of rabbits; decide to continue or terminate
|Pre-fusion booster||Day 62||Boost with 0.25 mg of antigen in saline, intraperitoneal|
|Fusion and ELISA screening||Day 76||Fuse myeloma cells and spleen cells, screen clones|
|ELISA and WB screening||Day 83||Screen clones|
|Subcloning and ELISA test||Day 97||Subclone to ensure monoclonal lines and ELISA test|
|Expansion and WB test||Day 104||
Expansion clones, then freeze stock;
Send supernatants to customer for validation
|Delivery||Day 110||Deliver the product customer choose and final report|