ArticleThe journal of histochemistry and cytochemistry : official journal of the Histochemistry Society2020
Proximity Ligation Assay as a Tool for Antibody Validation in Human Tissues.
Article in The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- Validation of an organ mapping antibody panel for cyclical immunofluorescence microscopy on normal human kidneys.American journal of physiology. Renal physiology · 2024Article
- Specific Detection of Physiological S129 Phosphorylated α-Synuclein in Tissue Using Proximity Ligation Assay.Journal of Parkinson's disease · 2023Article
- Let's not take DNA breaks for granted. The importance of direct detection of DNA breaks for the successful development of DDR inhibitors.Frontiers in cell and developmental biology · 2023Review
- Monitoring drug-target interactions through target engagement-mediated amplification on arrays and in situ.Nucleic acids research · 2022Article
- The Use of Quantitative Digital Pathology to Measure Proteoglycan and Glycosaminoglycan Expression and Accumulation in Healthy and Diseased Tissues.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2021Review
- Enhanced Validation of Antibodies Enables the Discovery of Missing Proteins.Journal of proteome research · 2020Article
Corrections and comments
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Authors and funding
7 authors.
Funding
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Abstract
Immunohistochemistry (IHC) is the accepted standard for spatial analysis of protein expression in tissues. IHC is widely used for cancer diagnostics and in basic research. The development of new antibodies to proteins with unknown expression patterns has created a demand for thorough validation. We have applied resources from the Human Protein Atlas project and the Antibody Portal at National Cancer Institute to generate protein expression data for 12 proteins across 39 cancer cell lines and 37 normal human tissue types. The outcome of IHC on consecutive sections from both cell and tissue microarrays using two independent antibodies for each protein was compared with in situ proximity ligation (isPLA), where binding by both antibodies is required to generate detection signals. Semi-quantitative scores from IHC and isPLA were compared with expression of the corresponding 12 transcripts across all cell lines and tissue types. Our results show a more consistent correlation between mRNA levels and isPLA as compared to IHC. The main benefits of isPLA include increased detection specificity and decreased unspecific staining compared to IHC. We conclude that implementing isPLA as a complement to IHC for analysis of protein expression and in antibody validation pipelines can lead to more accurate localization of proteins in tissue.
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