Control Line

Immunizing a host animal with the antibody(s) from a different species can generate secondary antibodies. For example, injecting mouse antibodies into an animal other than a mouse raise anti-mouse secondary antibodies. Goat, donkey, and rabbit are the most commonly used host species for producing secondary antibodies.

The most common types of secondary antibodies are those generated against a pooled population of immunoglobulins from a target species.

For example, immunizing a goat with purified mouse IgG will generate goat anti-mouse IgG antibodies that will bind to all classes, heavy and light chains (H+L) and fragments of mouse IgG as well as any other molecules sharing the strictly conserved domains (e.g., IgM share the same kappa light chains as IgG).

In contrast, immunizing a goat with only mouse IgG1 antibodies will generate antibodies specific to mouse IgG1 antibodies and molecules sharing the strictly conserved domains.

Because of the high degree of conservation in the structure of many immunoglobulin domains, class-specific secondary antibodies must be affinity purified and cross-adsorbed to achieve minimal cross-reaction with other immunoglobulins.

Immobilized mouse IgG1 antibodies can purify all goat antibodies that bind to mouse IgG1 using the above example. Passage through a chromatography column(s) containing mouse IgG2a, IgG2b, IgG3, IgM, etc., would further refine these anti-mouse IgG1 antibodies to remove any cross-react antibodies with non-IgG1 isotypes.

Additionally, passage through columns containing immobilized serum proteins from species other than those used to immunize the host can further purify secondary antibodies. This method of cross-adsorption (often referred to as "Highly Cross-Adsorbed") is an additional purification step recommended for applications when using primary antibodies from multiple species and when immunoglobulins or other serum proteins may be present in the samples probed.

Control Line Products

Antibody	Application
Rabbit IgG	For immunodiagnostic: ELISA, LFA, CLIA
Mouse IgG
Mouse IgM
Chicken IgY
Goat Anti Mouse IgG
Goat Anti Rabbit IgG
Goat Anti Chicken IgY
Mouse Anti Human IgG

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