To secure the success of experiments with antibodies, information provided about given catalogue antibody should contain the following: Antibody validation process is explained here. | Things to consider: Host Most popular host for a polyclonal antibody is rabbit. Antibodies produced in different hosts, offer various advantages. Clonality Polyclonal antibody is a mixture of antibodies recognizing various epitopes (from 3 to 15 amino acid long). Monoclonal and recombinant antibody binds to one epitope only. Antigen If a full length recombinant protein was used to produce an antibody to a protein which comes from a family with many isoforms, an antibody may react to all isoforms. Therefore, antibodies to short peptides are called monospecific, as they target a unique sequence within a target protein, chosen following in depth bioinformatic analysis. if short peptides are used fpr antibody production, one has to inquire antibody supplier about specific peptide location to make sure that correct antibody is going to be used on the sample from species of interest. Species reactivity Antibodies should not be used without checking that a given sequence used in its production is also found in the sequence which is aimed to be detected. This is often overlooked and leads to purchase and use of antibodies, which do not match target protein. Applications Antibody titer in ELISA is not representative for antibody reactivity on endogenous sample and can not be used as any guarantee. Application examples /Validation data Provided examples of antibody use, should also contain all detailes of applied protocol. If an antibody is detecting a recombinant protein, it is not a proof of its reactivity on endogenous sample, which has to be determined in a separate experiment. References As there is no common guideline on how antibodies should be cited in scientific publications, more than a half of antibodies used in research cannot be properly identified. |
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