Widjaja Anthony, Sidorova Julia M
Center for Genomics and Systems Biology , New York University, New York, New York, United States.
Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, United States.
MicroPubl Biol. 2025 Jan 31;2025. doi: 10.17912/micropub.biology.001485. eCollection 2025.
DNA fiber assays are indispensable tools for studying DNA damage and replication stress responses at the single replication fork level. These assays typically rely on antibodies recognizing IdU and CldU. Historically, the availability of CldU-reactive antibodies has been limited to one reagent (clone BU1/75(ICR1)). We validated two alternative antibodies for CldU detection in DNA fiber assays. One of these antibodies can be readily paired with a common IdU-reactive antibody, and we confirmed that it produces quantitatively similar CldU track length results vis-à-vis the BU1/75 antibody. The new reagents should boost versatility of DNA fiber assays, facilitating DNA replication research.
DNA纤维分析是在单复制叉水平研究DNA损伤和复制应激反应的不可或缺的工具。这些分析通常依赖于识别IdU和CldU的抗体。从历史上看，CldU反应性抗体的可用性仅限于一种试剂（克隆号BU1/75(ICR1)）。我们验证了两种用于DNA纤维分析中检测CldU的替代抗体。其中一种抗体可以很容易地与常见的IdU反应性抗体配对，并且我们证实，与BU1/75抗体相比，它产生的CldU轨道长度结果在数量上相似。这些新试剂应能提高DNA纤维分析的通用性，促进DNA复制研究。
