ArticleBiological procedures online2016
Fluorescence ImmunoPrecipitation (FLIP): a Novel Assay for High-Throughput IP.
Article in Biological procedures online, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
5 citing papers in PubMed.
- A Novel Confocal Scanning Protein-Protein Interaction Assay (PPI-CONA) Reveals Exceptional Selectivity and Specificity of CC0651, a Small Molecule Binding Enhancer of the Weak Interaction between the E2 Ubiquitin-Conjugating Enzyme CDC34A and Ubiquitin.Bioconjugate chemistry · 2024Article
- A polyclonal antibody against a recombinantly expressed Triticum aestivum RHT-D1A protein.Journal, genetic engineering & biotechnology · 2020Article
- LINE-1 ORF2p expression is nearly imperceptible in human cancers.Mobile DNA · 2020Article
- Article
- A toolbox of immunoprecipitation-grade monoclonal antibodies to human transcription factors.Nature methods · 2018Article
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
backgroundThe 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.
resultsHere 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.
conclusionsAltogether, our characterizations of this new methodology show that FLIP is an appealing and reliable tool for any application of high-throughput IP.
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Registered trials
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