We recommend using highly cross-adsorbed secondary antibodies whenever you perform multiple staining with two primary antibodies from different hosts. It is particularly important to use highly cross-adsorbed antibodies when staining with antibodies from two closely related species, such as mouse and rat.
Using highly cross-adsorbed secondary antibodies when staining tissues from a species with primary antibodies from a closely related species (for example, staining rat tissue with mouse primary antibody), will prevent background from cross-reactivity of the secondary with endogenous immunoglobulin in the tissue. This is important when staining immune tissues, but background from endogenous immunoglobulin is common in other tissues around blood vessels, where immunoglobulin also is abundant.
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