Evaluation of two new antibodies for recognition of CldU in DNA fiber assay applications.

microPublication biology Pub Date : 2025-01-31 eCollection Date: 2025-01-01 DOI:10.17912/micropub.biology.001485
Anthony Widjaja, Julia M Sidorova
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Abstract

DNA fiber assays are indispensable tools for studying DNA damage and replication stress responses in vivo 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.

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两种新的CldU识别抗体在DNA纤维分析中的应用评价。
DNA纤维分析是在单复制叉水平上研究体内DNA损伤和复制应激反应不可缺少的工具。这些检测通常依赖于识别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BU1/75(ICR1))。我们在DNA纤维分析中验证了两种用于CldU检测的替代抗体。其中一种抗体可以很容易地与一种常见的idu反应性抗体配对,我们证实它与-à-vis BU1/75抗体产生定量相似的CldU径迹长度结果。新试剂将提高DNA纤维分析的通用性,促进DNA复制研究。
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