Preparation of Polyclonal Antibodies to Retinoid Receptors

Techniques for the preparation of antibodies and their applications are widely established in the literature. Antibodies have proven to be useful reagents in characterizing the molecular activities of the retinoid receptors (1 ). The use of a retinoid “X” receptor α (RXRα) antibody on Western blots contributed to the identification of the RXR ligand by detection of the receptor in fractions of whole-cell lysates that were able to bind the 9-cis retinoic acid ligand (2 ) Antibodies to the retinoid receptors used in supershifting gelretardation assays also supported the findmg that RXR is the heterodimeric partner of other members of the retinoid/thyroid hormone-receptor superfamtly (3 , 4 ). Additionally, immunohistochemistry with antibodies to retinoic acid receptor a (RARα) enabled visualization of the promyelocyte-retinoic acid receptor (PML-RAR) fusion oncoprotein in cells from acute promyelocytrcleukemia patients (5 , 6 ) This chapter describes basic protocols for polyclonal rabbit-antibody production and affinity purification. Suggestions for potential applications in the study of retinoid-molecular biology are also discussed.