Assessing the Immune Response When Raising Antibodies for Use in Xenopus

Abstract

Frog-specific antibodies usually must be raised for work in Xenopus. Selecting a host animal whose immune system will respond to a target antigen with an antibody response is essential to obtaining high-quality antibodies. To determine whether an immunized animal has produced antibodies against an antigen, western blotting using Xenopus embryo or egg extract as the protein source can be performed as described here. When a protein of the expected size is detected by western blotting in the immune sera but not the preimmune sera, the antibody has been successfully raised.