BACKGROUND The immunoprecipitation (IP) assay is a valuable molecular biology tool applied across a breadth of fields. The standard assay couples IP to immunoblotting (IP/IB), a procedure severely limited as it is not easily scaled for high-throughput analysis. RESULTS Here we describe and characterize a new methodology for fast and reliable evaluation of an immunoprecipitation reaction. FLIP (FLuorescence IP) relies on the expression of the target protein as a chromophore-tagged protein and couples IP with the measurement of fluorescent signal coating agarose beads. We show here that FLIP displays similar sensitivity to the standard IP/IB procedure but is amenable to high-throughput analysis. We applied FLIP to the screening of mouse monoclonal antibodies of unknown behavior in IP procedures. The parallel analysis of the considered antibodies using FLIP and IP/western shows good correlation between the two procedures. We also show application of FLIP using unpurified antibodies (hybridoma supernatant) and we developed a publicly available tool for the easy analysis and quantification of FLIP signals. CONCLUSIONS Altogether, our characterizations of this new methodology show that FLIP is an appealing and reliable tool for any application of high-throughput IP.

中文翻译：

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荧光免疫沉淀 (FLIP)：一种新颖的高通量 IP 检测方法。

背景技术免疫沉淀（IP）测定是一种应用于广泛领域的有价值的分子生物学工具。标准检测将 IP 与免疫印迹 (IP/IB) 结合起来，这一过程受到严重限制，因为它不容易扩展到高通量分析。结果在这里，我们描述并描述了一种快速可靠评估免疫沉淀反应的新方法。FLIP（荧光 IP）依赖于目标蛋白作为发色团标记蛋白的表达，并将 IP 与琼脂糖珠涂层荧光信号的测量结合起来。我们在此表明​​，FLIP 显示出与标准 IP/IB 程序相似的灵敏度，但适合高通量分析。我们应用 FLIP 筛选 IP 程序中行为未知的小鼠单克隆抗体。使用 FLIP 和 IP/western 对所考虑的抗体进行平行分析，显示两种程序之间具有良好的相关性。我们还展示了使用未纯化的抗体（杂交瘤上清液）进行 FLIP 的应用，并且我们开发了一种公开可用的工具，用于轻松分析和量化 FLIP 信号。结论 总而言之，我们对这种新方法的描述表明，FLIP 对于任何高通量 IP 应用来说都是一个有吸引力且可靠的工具。