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Thus, it is essential for several pathways of repair of DNA damage by cisplatin and related drugs, which are widely used in the treatment of non\u2013small cell lung carcinoma and other late-stage tumors. Consequently, there is tremendous interest in measuring ERCC1-XPF expression in tumor samples. Many immunohistochemistry studies have been done, but the antibodies for ERCC1-XPF were not rigorously tested for antigen specificity. Herein, we survey a battery of antibodies raised against human ERCC1 or XPF for their specificity using ERCC1-XPF\u2013deficient cells as a negative control. Antibodies were tested for the following applications: immunoblotting, immunoprecipitation from cell extracts, immunofluorescence detection in fixed cells, colocalization of ERCC1-XPF with UV radiation\u2013induced DNA damage in fixed cells, and immunohistochemistry in paraffin-embedded samples. Although several commercially available antibodies are suitable for immunodetection of ERCC1-XPF in some applications, only a select subset is appropriate for detection of this repair complex in fixed specimens. The most commonly used antibody, 8F1, is not suitable for immunodetection in tissue. The results with validated antibodies reveal marked differences in ERCC1-XPF protein levels between samples and cell types. [Cancer Res 2009;69(17):6831\u20138]<\/jats:p>","DOI":"10.1158\/0008-5472.can-09-1237","type":"journal-article","created":{"date-parts":[[2009,9,1]],"date-time":"2009-09-01T00:34:23Z","timestamp":1251765263000},"page":"6831-6838","update-policy":"https:\/\/doi.org\/10.1158\/crossmark_policy","source":"Crossref","is-referenced-by-count":86,"title":["Immunodetection of DNA Repair Endonuclease ERCC1-XPF in Human Tissue"],"prefix":"10.1158","volume":"69","author":[{"given":"Nikhil R.","family":"Bhagwat","sequence":"first","affiliation":[{"name":"1Department of Human Genetics, University of Pittsburgh School of Public Health;"},{"name":"2University of Pittsburgh Cancer Institute, Hillman Cancer Center; Departments of"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vera Y.","family":"Roginskaya","sequence":"additional","affiliation":[{"name":"2University of Pittsburgh Cancer Institute, Hillman Cancer Center; Departments of"},{"name":"3Pharmacology and Chemical Biology and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie B.","family":"Acquafondata","sequence":"additional","affiliation":[{"name":"5Department of Pathology, University of Pittsburgh Medical Center Shadyside, Pittsburgh, Pennsylvania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rajiv","family":"Dhir","sequence":"additional","affiliation":[{"name":"5Department of Pathology, University of Pittsburgh Medical Center Shadyside, Pittsburgh, Pennsylvania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard D.","family":"Wood","sequence":"additional","affiliation":[{"name":"2University of Pittsburgh Cancer Institute, Hillman Cancer Center; Departments of"},{"name":"3Pharmacology and Chemical Biology and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura J.","family":"Niedernhofer","sequence":"additional","affiliation":[{"name":"2University of Pittsburgh Cancer Institute, Hillman Cancer Center; Departments of"},{"name":"4Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine;"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1086","published-online":{"date-parts":[[2009,8,31]]},"reference":[{"key":"2022061701482605600_B1","unstructured":"In: Ries LAG MD, Krapcho M, Stinchcomb DG, et al., editors. 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