A simple treatment to significantly increase signal specificity in immunohistochemistry

It is increasingly important to understand a gene’s function at the protein level. Immunohistochemistry is used to identify new functions of phytohormones by finding their precise localization patterns. To facilitate protein immunolocalization, we developed a protocol that can significantly reduce background and give highly specific signals of antibody to traget proteins. The key improvement is to treat secondary antibodies by mixing with homogenized fresh and fixed plant tissue. Using this protocol, we have successfully carried out immunolocalization of proteins encoded by genes AP3 and HoMADS2 , as well as plant hormones ABA and IAA.