Research Antibodies

Primary and secondary antibodies differ in how they bind to antigens.

Primary antibodies bind directly to antigens in a tissue or sample, whereas secondary antibodies bind to primary antibodies creating an antigen-primary-secondary chain.

Direct immunoassays use primary antibodies to detect antigens. Indirect immunoassays use both primary and secondary antibodies.

An isotype control is a primary antibody that lacks specificity against the target antigen but matches the type and class of the primary antibody used in the experiment.

They serve as negative controls that generate reliable data by allowing researchers to accurately differentiate between specific antibody signal and non-specific interactions causing background signal.

Antibodies for flow cytometry allow scientists to detect specific antigens and characterize the proteins on the surface of cells. Similar to other immunoassays, antibodies used in flow cytometry can be directly labeled with a fluorescent dye or bind to a secondary antibody carrying the fluorescent marker. The specialized nature of antibodies enables flow cytometry to detect different cell types according to their fluorescent color, each representing an antigen-antibody interaction.

Antibodies are created by inoculating an animal with an antigen and harvesting antibodies from blood serum.

Choose a primary antibody that was produced in a host species that is different from the species that provided your sample. For example, if you are studying mouse samples, choose a primary that was not raised in mouse.

Find a secondary antibody that is against the host species of your primary antibody. For example, if the primary antibody was raised in mouse, choose an anti-mouse secondary.