ArticleNature protocols2013
Solid-phase proximity ligation assays for individual or parallel protein analyses with readout via real-time PCR or sequencing.
Article in Nature protocols, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 23 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
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.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
23 citing papers in PubMed, 58 citations in OpenAlex.
- Proteomics-The State of the Field: The Definition and Analysis of Proteomes Should Be Based in Reality, Not Convenience.Proteomes · 2024Article
- A streamlined proximity extension assay using POEGMA polymer-coated magnetic beads for enhanced protein detection.Frontiers in bioengineering and biotechnology · 2024Article
- Extending the dynamic range of biomarker quantification through molecular equalization.Nature communications · 2023Article
- Assay methods based on proximity-enhanced reactions for detecting non-nucleic acid molecules.Frontiers in bioengineering and biotechnology · 2023Review
- Surface protein profiling of prostate-derived extracellular vesicles by mass spectrometry and proximity assays.Communications biology · 2022Article
- Article
- One-pot pre-coated interface proximity extension assay for ultrasensitive co-detection of anti-SARS-CoV-2 antibodies and viral RNA.Biosensors & bioelectronics · 2021Article
- Proximity ligation assay: an ultrasensitive method for protein quantification and its applications in pathogen detection.Applied microbiology and biotechnology · 2021Review
- Re-Evaluating the Conventional Wisdom about Binding Assays.Trends in biochemical sciences · 2020Review
- Ultrasensitive digital quantification of cytokines and bacteria predicts septic shock outcomes.Nature communications · 2020Article
- A Concise, Modular Antibody-Oligonucleotide Conjugation Strategy Based on Disuccinimidyl Ester Activation Chemistry.Chembiochem : a European journal of chemical biology · 2019Article
- Nucleic Acid-Barcoding Technologies: Converting DNA Sequencing into a Broad-Spectrum Molecular Counter.Angewandte Chemie (International ed. in English) · 2019Review
- Proximity Ligation Assay (PLA) to Determine the Endosomal Localization of ESCRT Subunit in Virus-Infected Cells.Methods in molecular biology (Clifton, N.J.) · 2019Article
- Antibody validation: a view from the mountains.New biotechnology · 2018Review
- Nanoparticle-Based Proximity Ligation Assay for Ultrasensitive, Quantitative Detection of Protein Biomarkers.ACS applied materials & interfaces · 2018Article
- Sensitive and Specific Detection of Platelet-Derived and Tissue Factor-Positive Extracellular Vesicles in Plasma Using Solid-Phase Proximity Ligation Assay.TH open : companion journal to thrombosis and haemostasis · 2018Article
- Evaluation of a New Survivin ELISA and UBCInternational journal of molecular sciences · 2018Article
- Evaluation of circulating cellular DCLK1 protein, as the most promising colorectal cancer stem cell marker, using immunoassay based methods.Cancer biomarkers : section A of Disease markers · 2016Article
- Detection of Biomarkers with Solid-Phase Proximity Ligation Assay in Patients with Colorectal Cancer.Translational oncology · 2016Article
- Proximity assays for sensitive quantification of proteins.Biomolecular detection and quantification · 2015Review
Corrections and comments
- Erratum issued
Authors and funding
8 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Solid-phase proximity ligation assays share properties with the classical sandwich immunoassays for protein detection. The proteins captured via antibodies on solid supports are, however, detected not by single antibodies with detectable functions, but by pairs of antibodies with attached DNA strands. Upon recognition by these sets of three antibodies, pairs of DNA strands brought in proximity are joined by ligation. The ligated reporter DNA strands are then detected via methods such as real-time PCR or next-generation sequencing (NGS). We describe how to construct assays that can offer improved detection specificity by virtue of recognition by three antibodies, as well as enhanced sensitivity owing to reduced background and amplified detection. Finally, we also illustrate how the assays can be applied for parallel detection of proteins, taking advantage of the oligonucleotide ligation step to avoid background problems that might arise with multiplexing. The protocol for the singleplex solid-phase proximity ligation assay takes ~5 h. The multiplex version of the assay takes 7-8 h depending on whether quantitative PCR (qPCR) or sequencing is used as the readout. The time for the sequencing-based protocol includes the library preparation but not the actual sequencing, as times may vary based on the choice of sequencing platform.
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.