Sensitive Method for Determination of Protein and Cell Concentration Based on Competitive Adsorption to Nanoparticle and Time-Resolved Luminescence Resonance Energy Transfer between Labeled Proteins.
Summary of "Sensitive Method for Determination of Protein and Cell Concentration Based on Competitive Adsorption to Nanoparticle and Time-Resolved Luminescence Resonance Energy Transfer between Labeled Proteins."
A sensitive mix-and-measure method for the determination of protein and cell concentration was developed. It is based on the competitive adsorption between the analyte and donor- and acceptor-labeled proteins to carboxylate-modified polystyrene nanoparticles. High time-resolved luminescence resonance energy transfer (TR-LRET) signal is detected in the absence of the analyte due to the close proximity of the nanoparticle-adsorbed labeled proteins. The increased concentration of the analyte decreases the adsorption of the labeled proteins leading to the loss of proximity and thus, decrease in the TR-LRET. The detection limit of the assay was 2.6 ng of proteins, which is higher than that of the most sensitive commercial methods. The method was also applied to cell counting and 200 eukaryotic cells were measured in a microtiter well under the optimized conditions. The average coefficient of variation for both developed assays was approximately 10% and the protein-to-protein variability for eleven different proteins was no more than 20%. The developed method requires no labeled particles, making the concept optimally applicable to varying targets as the material of the particle may be selected according to the application.
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Name: Analytical chemistry
Medical and Biotech [MESH] Definitions
Blood Volume Determination
Method for determining the circulating blood volume by introducing a known quantity of foreign substance into the blood and determining its concentration some minutes later when thorough mixing has occurred. From these two values the blood volume can be calculated by dividing the quantity of injected material by its concentration in the blood at the time of uniform mixing. Generally expressed as cubic centimeters or liters per kilogram of body weight.
Autocrine Motility Factor
A protein cytokine secreted by tumor cells. It elicits increases in cell motility and phosphoinositide metabolism in the secreting cell via a pertussis toxin-sensitive G-protein signal transduction pathway. It is closely related to PHOSPHOHEXOSE ISOMERASE; NEUROLEUKIN; and maturation factor.
Gtp-binding Protein Alpha Subunit, Gi2
A PERTUSSIS TOXIN-sensitive GTP-binding protein alpha subunit. It couples with a variety of CELL SURFACE RECEPTORS, has been implicated in INTERLEUKIN-12 production, and may play a role in INFLAMMATORY BOWEL DISEASES.
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SNARE binding proteins that facilitate the ATP hydrolysis-driven dissociation of the SNARE complex. They are required for the binding of N-ETHYLMALEIMIDE-SENSITIVE PROTEIN (NSF) to the SNARE complex which also stimulates the ATPASE activity of NSF. They are unrelated structurally to SNAP-25 PROTEIN.
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Visible morphologic changes in cells infected with viruses. It includes shutdown of cellular RNA and protein synthesis, cell fusion, release of lysosomal enzymes, changes in cell membrane permeability, diffuse changes in intracellular structures, presence of viral inclusion bodies, and chromosomal aberrations. It excludes malignant transformation, which is CELL TRANSFORMATION, VIRAL. Viral cytopathogenic effects provide a valuable method for identifying and classifying the infecting viruses.
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