ArticleCytometry. Part A : the journal of the International Society for Analytical Cytology2010
Tyramide signal amplification for analysis of kinase activity by intracellular flow cytometry.
Article in Cytometry. Part A : the journal of the International Society for Analytical Cytology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 45 citations in OpenAlex.
- Assessment and detection of nucleotide excision repair capability in human cells: a critical review of current functional methods.Molecular biology reports · 2026Review
- TyrRu: A Tyramide-Amplified Multimodal Ruthenium Tag for Elemental Mass Spectrometry and Immunofluorescence Imaging.Chemical & biomedical imaging · 2026Article
- Catch-and-display immunoassay for digital biomarker detection.Nanoscale horizons · 2026Article
- A Customizable Tyramide Signal Amplification-Based Multiplex Immunofluorescence Protocol for FFPE Tissues.Current issues in molecular biology · 2026Article
- Comprehensive Overview of Gastric Cancer Immunohistochemistry: Key Biomarkers, Advanced Detection Methods, and Perspectives.Medicina (Kaunas, Lithuania) · 2026Review
- Antibody-Based Multiplex Image Analysis: Standard Analytical Workflows and Artificial Intelligence Tools for Pathologists.Laboratory investigation; a journal of technical methods and pathology · 2025Review
- Article
- Magnetic Silica-Coated Fluorescent Microspheres (MagSiGlow) for Simultaneous Detection of Tumor-Associated Proteins.Angewandte Chemie (International ed. in English) · 2024Article
- Type-Independent 3D Writing and Nano-Patterning of Confined Biopolymers.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2023Article
- High-throughput screening method for discovering CatSper inhibitors using membrane depolarization caused by external calcium chelation and fluorescent cell barcoding.Frontiers in cell and developmental biology · 2023Article
- Automated quantitative high-throughput multiplex immunofluorescence pipeline to evaluate OXPHOS defects in formalin-fixed human prostate tissue.Scientific reports · 2022Article
- A Hybrid Detection Method Based on Peroxidase-mediated Signal Amplification and Click Chemistry for Highly Sensitive Background-free Immunofluorescent Staining.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2019Article
- Immuno-SABER enables highly multiplexed and amplified protein imaging in tissues.Nature biotechnology · 2019Article
- Amplification of unscheduled DNA synthesis signal enables fluorescence-based single cell quantification of transcription-coupled nucleotide excision repair.Nucleic acids research · 2017Article
- Bright Fluorescent Nanotags from Bottlebrush Polymers with DNA-Tipped Bristles.ACS central science · 2015Article
- Multiparameter flow cytometry: advances in high resolution analysis.Immune network · 2013Article
- Glutathione S-transferase P1 correlated with oxidative stress in hepatocellular carcinoma.International journal of medical sciences · 2013Article
- Spectral flow cytometry.Current protocols in cytometry · 2013Review
- Single-cell protein analysis.Current opinion in biotechnology · 2012Review
- Novel hematopoietic progenitor populations revealed by direct assessment of GATA1 protein expression and cMPL signaling events.Stem cells (Dayton, Ohio) · 2011Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
Intracellular flow cytometry permits quantitation of diverse molecular targets at the single-cell level. However, limitations in detection sensitivity inherently restrict the method, sometimes resulting in the inability to measure proteins of very low abundance or to differentiate cells expressing subtly different protein concentrations. To improve these measurements, an enzymatic amplification approach called tyramide signal amplification (TSA) was optimized for assessment of intracellular kinase cascades. First, Pacific Blue, Pacific Orange, and Alexa Fluor 488 tyramide reporters were shown to exhibit low nonspecific binding in permeabilized cells. Next, the effects of antibody concentration, tyramide concentration, and reaction time on assay resolution were characterized. Use of optimized TSA resulted in a 10-fold or greater improvement in measurement resolution of endogenous Erk and Stat cell signaling pathways relative to standard, nonamplified detection. TSA also enhanced assay sensitivity and, in conjunction with fluorescent cell barcoding, improved assay performance according to a metric used to evaluate high-throughput drug screens. TSA was used to profile Stat1 phosphorylation in primary immune system cells, which revealed heterogeneity in various populations, including CD4+ FoxP3+ regulatory T cells. We anticipate the approach will be broadly applicable to intracellular flow cytometry assays with low signal-to-noise ratios.
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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.