ArticleAnalytical biochemistry2015
Quantitation of secreted proteins using mCherry fusion constructs and a fluorescent microplate reader.
Article in Analytical biochemistry, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 24 citations in OpenAlex.
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- Beyond the surface: the plant secretome as a bridge between the cell and its environment.Planta · 2025Review
- snoRNA-facilitated protein secretion revealed by transcriptome-wide snoRNA target identification.Cell · 2025Article
- Oriented Insertion of ESR-Containing Hybrid Proteins in Proteoliposomes.International journal of molecular sciences · 2023Article
- Peroxisome proliferator-activated receptor gamma-ligand-binding domain mutations associated with familial partial lipodystrophy type 3 disrupt human trophoblast fusion and fibroblast migration.Journal of cellular and molecular medicine · 2020Article
- A novel system to map protein interactions reveals evolutionarily conserved immune evasion pathways on transmissible cancers.Science advances · 2020Article
- mCherry fusions enable the subcellular localization of periplasmic and cytoplasmic proteins in Xanthomonas sp.PloS one · 2020Article
- Nucleic acid-induced potentiation of matrix metalloproteinase-9 enzymatic activity.The Biochemical journal · 2018Article
- Neonicotinoid-induced pathogen susceptibility is mitigated by Lactobacillus plantarum immune stimulation in a Drosophila melanogaster model.Scientific reports · 2017Article
- Secreted dual reporter assay with Gaussia luciferase and the red fluorescent protein mCherry.PloS one · 2017Article
- Functional Roles of N-Linked Glycosylation of Human Matrix Metalloproteinase 9.Traffic (Copenhagen, Denmark) · 2015Article
- LMAN1 (ERGIC-53) is a potential carrier protein for matrix metalloproteinase-9 glycoprotein secretion.Biochemical and biophysical research communications · 2015Article
- Nanoparticle-Based and Bioengineered Probes and Sensors to Detect Physiological and Pathological Biomarkers in Neural Cells.Frontiers in neuroscience · 2015Review
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Traditional assays for secreted proteins include methods such as Western blot and enzyme-linked immunosorbent assay (ELISA) detection of the protein in the cell culture medium. We describe a method for the detection of a secreted protein based on fluorescent measurement of an mCherry fusion reporter. This microplate reader-based mCherry fluorescence detection method has a wide dynamic range of 4.5 orders of magnitude and a sensitivity that allows detection of 1 to 2fmol fusion protein. Comparison with the Western blot detection method indicated greater linearity, wider dynamic range, and a similar lower detection threshold for the microplate-based fluorescent detection assay of secreted fusion proteins. An mCherry fusion protein of matrix metalloproteinase-9 (MMP-9), a secreted glycoprotein, was created and expressed by transfection of human embryonic kidney (HEK) 293 cells. The cell culture medium was assayed for the presence of the fluorescent signal up to 32 h after transfection. The secreted MMP-9-mCherry fusion protein was detected 6h after transfection with a linear increase in signal intensity over time. Treatment with chloroquine, a drug known to inhibit the secretion of many proteins, abolished the MMP-9-mCherry secretion, demonstrating the utility of this method in a biological experiment.
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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.