ArticleLaboratory investigation; a journal of technical methods and pathology2017
Proof of the quantitative potential of immunofluorescence by mass spectrometry.
Article in Laboratory investigation; a journal of technical methods and pathology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed, 43 citations in OpenAlex.
- A semi-automated pipeline integrating ImageJ/Fiji and StarDist for the reproducible quantification of cellular and optical density metrics in immunofluorescence images of brain tissue.Frontiers in neuroanatomy · 2026Article
- A PQQ-dependent oxidoreductase inFrontiers in cellular and infection microbiology · 2026Article
- Immunofluorescent Quantification of MIF in Larval Zebrafish Tailfin Transection.Methods in molecular biology (Clifton, N.J.) · 2026Article
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- Quantitative Protein Expression of Antibody-Drug Conjugate Targets in EGFR Mutated and Wild-type Non-Small Cell Lung Cancer.Clinical cancer research : an official journal of the American Association for Cancer Research · 2025Article
- Transcriptomic Approaches to Cardiomyocyte-Biomaterial Interactions: A Review.ACS biomaterials science & engineering · 2024Review
- Automated identification of protein expression intensity and classification of protein cellular locations in mouse brain regions from immunofluorescence images.Medical & biological engineering & computing · 2024Article
- High fat, high sucrose diet promotes increased expression of ACE2 receptor in the SIV-infected host: implications for SARS-CoV-2 infection.Frontiers in nutrition · 2024Article
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- Rapid responses of human pluripotent stem cells to cyclic mechanical strains applied to integrin by acoustic tweezing cytometry.Scientific reports · 2023Article
- Digital spatial profiling of melanoma shows CD95 expression in immune cells is associated with resistance to immunotherapy.Oncoimmunology · 2023Article
- A comparative study of gastric adenocarcinoma HER2 IHC phenotype and mass spectrometry-based quantification.Frontiers in oncology · 2023Article
- Deep learning tools and modeling to estimate the temporal expression of cell cycle proteins from 2D still images.PLoS computational biology · 2022Article
- Cancer microenvironment and genomics: evolution in process.Clinical & experimental metastasis · 2022Review
- Improved Methods for Single-Molecule Fluorescence In Situ Hybridization and Immunofluorescence in Caenorhabditis elegans Embryos.Current protocols · 2021Article
- Best Practices for Technical Reproducibility Assessment of Multiplex Immunofluorescence.Frontiers in molecular biosciences · 2021Review
- Antibody validation for protein expression on tissue slides: a protocol for immunohistochemistry.BioTechniques · 2020Article
- Dexamethasone and Glucocorticoid-Induced Matrix Temporally Modulate Key Integrins, Caveolins, Contractility, and Stiffness in Human Trabecular Meshwork Cells.Investigative ophthalmology & visual science · 2020Article
- Sensitive quantification of α-glucans in mouse tissues, cell cultures, and human cerebrospinal fluid.The Journal of biological chemistry · 2020Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 2 countries.
Funding
Abstract
Protein expression in formalin-fixed, paraffin-embedded patient tissue is routinely measured by Immunohistochemistry (IHC). However, IHC has been shown to be subject to variability in sensitivity, specificity and reproducibility, and is generally, at best, considered semi-quantitative. Mass spectrometry (MS) is considered by many to be the criterion standard for protein measurement, offering high sensitivity, specificity, and objective molecular quantification. Here, we seek to show that quantitative immunofluorescence (QIF) with standardization can achieve quantitative results comparable to MS. Epidermal growth factor receptor (EGFR) was measured by quantitative immunofluorescence in 15 cell lines with a wide range of EGFR expression, using different primary antibody concentrations, including the optimal signal-to-noise concentration after quantitative titration. QIF target measurement was then compared to the absolute EGFR concentration measured by Liquid Tissue-selected reaction monitoring mass spectrometry. The best agreement between the two assays was found when the EGFR primary antibody was used at the optimal signal-to-noise concentration, revealing a strong linear regression (R
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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.