Immunoaffinity purification of the human multisubunit transcription factor IIH

Abstract

A procedure to immunoaffinity purify the human transcription factor IIH (TFIIH) was developed using a monoclonal antibody that recognizes an epitope in ERCC3 (XPB), the largest subunit of TFIIH. The epitope recognized by the monoclonal antibody was mapped to 20 amino acids. A peptide containing the epitope was capable of displacing TFIIH from an immunoaffinity column containing the monoclconal antibody. The immunoaffinity purification procedure described allows a simple and efficient method to purify both the 'core' and 'holo' TFIIH complexes.