Immunolocalization of IAA Using an Anti-IAA-C-Antibody Raised Against Carboxyl-Linked IAA.

Plant hormone indole-3-acetic acid (IAA) plays a crucial role in plant physiological events such as plant development, differentiation, and environmental responses. IAA is synthesized in specific focal cells and/or tissues such as the coleoptile tip in maize and the root tip and young leaf primordia in Arabidopsis thaliana. Recent studies have shown that formation of an IAA maxima or concentration gradient, created by the changing expression and cellular localization of IAA transport proteins, crucially controls plant physiological events. For this reason, visualization of IAA molecules at the cell and tissue levels is necessary to accurately determine the distribution of IAA in plants. Immunolocalization of IAA is a means to directly visualize IAA and observe its localization and distribution in plant cells and tissues. Here, we introduce an immunolocalization protocol to observe IAA distribution that uses a specific anti-IAA-C-antibody raised against carboxyl-linked IAA. This method is applicable for various plant samples and is reliable for specifically detecting IAA in plant tissues.