Accelerate Biological Research
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+1-888.733.6849
+1-617.299.7367 (Int’l)
+1-617.299.7367 (Int’l)
+1-888.733.6849
+1-617.299.7367 (Int’l)
+1-617.299.7367 (Int’l)
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service@kendallscientific.com
+1-888.733.6849 (Toll-free)
+1-617.299.7367 (Int’l))
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Peptide synthesis: Phospho-amino acid modified peptide (>15 aa >90% purity, 15 mg) and unphosphorylated control (>15 aa >90% purity, 11 mg)
Animal immunization: 2 rabbits, multiple production bleeds
Phospho-specific ELISA
Affinity purification with phosphopeptide and cross-adsorption with non-phospho peptide
Lyophilization (optional)
1-30 mg purified antibody
5 mg phophopeptide
5 mg non-phospho peptide
1-2 ml of pre-immune serum
ELISA results, MS and HPLC reports
Peptide synthesis: Phospho-amino acid modified peptide (>15 aa >90% purity, 10 mg) and unphosphorylated control (>15 aa >90% purity, 6 mg)
Animal immunization: 10 mice or 6 rats
Test bleed ELISA and hybridoma screening for phophopeptide, non-phosphopeptide and generic phosphopeptide
Subcloning, expansion, cryopreservation of positive cell lines
Lyophilization of supernatants
2 positive hybridoma clones
5 ml positive supernatant for each clone
5 mg phophopeptide
5 mg non-phospho peptide
Kendall Scientific's standard option for polyclonal antibody is optimized to produce antiserum in the shortest possible time. It is the most affordable option, popular among researchers.
The protocol uses two New Zealand White rabbits that are Specific Pathogen-Free. Injections are subcutaneous (SQ) as emulsions in Complete Freund's Adjuvant (CFA) or Incomplete Freund's Adjuvant (IFA).
| Procedure | Protocol Day | Description |
|---|---|---|
| Control Serum Collection | Day 0 | Pre-immune bleed (5mL per rabbit) |
| Primary Injection | Day 1 | Immunize with 0.50mg of antigen in CFA, 10 SQ sites |
| 1st Booster | Day 14 | Boost with 0.25mg of antigen in IFA, 4 SQ sites |
| 2nd Booster | Day 28 | Boost with 0.25mg of antigen in IFA, 4 SQ sites |
| Serum Collection | Day 35 | Bleed (~25mL per rabbit) |
| 3rd Booster | Day 42 | Boost with 0.25mg of antigen in IFA, 4 SQ sites |
| Serum Collection | Day 56, 58 | Two bleeds (~50mL total per rabbit) |
| ELISA Test | Day 60 | ELISA titration (results available online); Verify disposition of rabbits; decide to continue or terminate |
| Purification | Optional | 6-10 mg purified antibodies |
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