Polyclonal Antibodies against Synthetic phospho-peptide

Phosphorylation is the addition of a phosphate (PO4) group to a protein molecule or a small molecule. Its prominent role in biochemistry is the subject of a very large body of research. Phosphorylation usually occurs on serine, threonine, and tyrosine residues in eukaryotic proteins. Antibodies can be used as powerful tools to detect whether a protein is phosphorylated at a particular site. Antibodies bind to and detect phosphorylation-induced conformational changes in the protein. Such antibodies are called phospho-specific antibodies. They are becoming critical reagents both for basic research and for clinical diagnosis.

Phospho-specific Antibodies are generated using, as antigen, a peptide containing a phosphorylated aminoacid residue, Primm helps you to select the best peptide sequence and suggests to use:
Short peptide
Phosphorylated aminoacid residue either at N- or C-terminus
Cys residue added for selected carrier conjugation

Primm Phospho Specific Polyclonal Antibody production includes:
Two rabbits immunization with a phospho-synthetic peptide conjugated either to OVA or KLH, Synthesis of the corresponding non phospho peptide for preparation of depletion column and for differential ELISA test,
Standard immunization schedule (5 injections - about 1.5/2 months),
Final bleed (40-50 ml/rabbit)
Serum positivity is guaranteed at a dilution of the serum equivalent to 1:1000 as compared to preimmune serum in differential ELISA test

Total immuno absorbtion of the immune sera using CNBr-Sepharose Non-phospho peptide column.
Affinity chromatography using CNBr-Sepharose phospho peptide column (keeping separated each purified prep of each rabbit).

Final poduct: Purified antibody specific for phospho-peptide.

Delivered products:
- 2 different aliquots of immunopurified phospho specific polyclonal antibody (+ ELISA test profile)
- pre-immune sera, 2 ml/rabbit
- free phospho peptide at least 2 mg
- free non-phospho peptide at least 2 mg
- affinity column (CNBr-Sepharose + phospho peptide)
- depletion column (CNBR-Sepharose + non-phospho peptide)

Time required:
3 months (3/4 weeks to synthesize peptides, 1.5/2 months to immunize animals and 1/2 weeks for immunoabsorption and immunoaffinity purification)

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