Peptides as Immunohistochemistry Controls

Protein Molecular Data from Ancient (>1 million years old) Fossil Material: Pitfalls, Possibilities and Grand Challenges

“…Each immunological assay is performed in conjunction with specificity tests to account for spurious binding. These include the following: (1) inhibition (also called “preadsorption”) assays, in which the paratopes present in an antibody are blocked with its purported immunogen before incubation with the sample, allowing the identification of additional paratopes contained in our polyclonal antibodies to molecules other than the target protein; − (2) digestion assays, in which the target protein is selectively destroyed (either partially or completely) before incubation with antibodies raised against it, to assess whether antigen–antibody complex formation is directly affected by the specific digestion of the target protein; (3) irrelevant antibody assays, in which samples are incubated with antibodies raised against proteins that are not typically found in the sample tissue (e.g., mammalian antitestosterone antibodies with dinosaur bone tissue) to confirm that the reactions to sample material are not indiscriminate . These are in addition to standard “secondary-only” assays included in every trial of every experiment, where a subset of samples are incubated with antibody buffer (lacking primary antibody) during the primary antibody step, then treated to secondary antibody and all other steps for signal detection to determine if the secondary may be binding nonspecifically to molecules in the tissue, and not exclusively to bound primary antigen–antibody complexes.…”