I am studying a tyrosine phosphorylation event on a protein and have both polyclonal and monoclonal antibodies that recognize the phosphorylated residue. My understanding is that polyclonal antibodies are comprised of antibodies that recognize different epitopes on a given antigen, while monoclonal antibodies recognize the same epitope on a given antigen. If a polyclonal antibody is created using a peptide sequence containing a phosphorylated amino acid, does this mean that the antibody can recognize some epitopes that do and some epitopes that do not (say, if the epitope is upstream of the phosphorylated residue) contain the phosphorylated residue? How are polyclonal antibodies phospho-specific? It would appear to me that only a monoclonal antibody with an epitope containing the phosphorylated tyrosine would be phospho-specific.
Hi, you are right, but when you sell or use a phospho specific polyclonal Ab, you would absorb non-phospho-specific Ab, for example by affinity chromatography against the non-phosphorylated sequence or Protein. After that, you still have a variety of Ab so they are polyclonal AND phospho-specific.
Hi Cody, polyclonal / monoclonal describes the clonality of the AB producing cells and doesn't include any information about the number of epitopes an AB preparation will bind to.
Example: If you immunize a rabbit, chicken or mouse with phosphotyrosine coupled to a carrier, you will get polyclonal sera which will recognize the carrier and hopefully the target (pTyr in your case). Then, when the antiserum is affinity purified, you will extract the antibodies which bind your target.
If you should decide to generate a mouse hybridoma, you will screen many clones and usually only keep the "best" one, but nevertheless, you'll start with an amount of different clones recognizing your target.
One can generate phospho-specific polyclonal antibody provided phospho epitope of antigen bind with B cell receptor with high affinity as compare to other epitope. Because of this antibody generation will start against phospho epitope of antigen first.
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