Ihc antibody cross reactivity8/25/2023 the composition of the individual subclasses and parts corresponds to the normal distribution in the serum of the species.Īntiserum from animals immunized with IgG(H+L) contains antibodies directed against both the light and heavy chains of the IgG molecule. As a rule, the immunoglobulins used for immunization are obtained from total serum, i.e. goat) in which the antiserum is made is immunized with purified immunoglobulin or immunoglobulin parts of the corresponding target species. Generation of an antibody with a specificity mouse IgG (H+L) In this case the optimal specificity for each of the secondary antibodies is still IgG(H+L). ![]() This is done by adsorption of antibodies. In this case, cross-recognition must be prevented. If only one primary antibody is to be detected in a planned assay and immunoglobulins of other classes or subclasses are not a confounding factor, as is the case in most applications, specificity IgG (H+L) is usually most appropriate.įrequently, several primary antibodies from different species are to be detected simultaneously in one experiment (multiple labeling). This reagent recognizes a variety of epitopes on antibodies of different classes and subclasses. Secondary antibodies should have a high sensitivity for this primary antibody.Īs a rule, this is a secondary antibody for which a whole molecule IgG(H+L) was used as the immunogen. from rabbit, goat, chicken, etc.) or monoclonal antibodies from mouse, rat or hamster, which are obtained from B-cell hybridomas, are used. ![]() Step 4: Antibody Specificity What should the secondary antibody detect?įor immunological assays, mainly polyclonal antibodies of the IgG type (e.g.
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