Book ChapterDOI
Quantitation of protein.
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TLDR
This chapter discusses various methods of estimating protein concentration as defined by the difference in energy between the orbital of the unexcited electron and a higher energy orbital.Abstract:
Publisher Summary This chapter discusses various methods of estimating protein concentration. Absorption spectroscopy involves the absorption of a photon by an electron. Only those photons with a certain energy level can be absorbed as defined by the difference in energy between the orbital of the unexcited electron and a higher energy orbital. The peptide bond absorbs photons below 210 nm. Because of the large number of peptide bonds in a protein, this is a highly sensitive area of the protein spectrum. Although protein conformation and some absorption by tryptophan and tyrosine residues occurs in this region, less variability between proteins is observed than at 280 nm. There is a need to avoid storing buffers in plastic containers because some plastics leach plasticizers, which absorb at ultraviolet (UV) wavelengths. Detergents can also be troublesome because many absorb UV light. If the buffer or protein solution is cold, the outside of the cuvette may need to be wiped between each reading with a lint-free wiper and the readings should be made quickly after placing the cold solution into the cuvette, because atmospheric moisture may condense on the outside of the cuvette producing an erroneously high reading.read more
Citations
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Journal ArticleDOI
N-Terminal Region of GbIspH1, Ginkgo biloba IspH Type 1, May Be Involved in the pH-Dependent Regulation of Enzyme Activity.
TL;DR: It was suggested that the N-terminal region of GbIspH1 is involved in the pH-dependent regulation of enzyme activity during photosynthesis.
Journal ArticleDOI
Stereoselective synthesis of conformationally restricted KOR agonists based on the 2,5-diazabicyclo[2.2.2]octane scaffold
TL;DR: 2,5-diazabicyclooctanes bearing a pyrrolidino moiety in the 7-position were envisaged to study KOR agonists with a conformationally rigid ethylenediamine pharmacophore and thus a defined N(pyrrolidine)-C7-C1-N2 dihedral angle.
Journal ArticleDOI
Quantitation of Nucleic Acids and Proteins
TL;DR: Two methods for direct absorbance measurements at 260 nm are described—the first is a traditional cuvette‐based method, and the second is a microvolume method that requires no cuvettes or capillaries.
Journal ArticleDOI
Discovery of new potent dual sigma receptor/GluN2b ligands with antioxidant property as neuroprotective agents
Daniele Zampieri,Sara Fortuna,Antonella Calabretti,Maurizio Romano,Renzo Menegazzi,Dirk Schepmann,Bernhard Wünsch,Simona Collina,Davide Zanon,Maria Grazia Mamolo +9 more
TL;DR: Results proved that the dual σR/GluN2b ligands have beneficial effects in a model of neuronal oxidative stress and can represent strong candidate pharmacotherapeutic agents for minimizing oxidative stress-induced neuronal injuries.
Dissertation
Production of poly(3-hydroxybutyrate) and ectoines using a halophilic bacterium
TL;DR: The present study has demonstrated the protective effect of ectoine and hydroxyectoine on enzymes with respect to pH stress, and it is suggested that the presence of ectoines results in reduced conformational flexibility of the biomolecules.
References
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Journal Article
Protein Measurement with the Folin Phenol Reagent
TL;DR: Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein.
Journal ArticleDOI
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
TL;DR: This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose.
PatentDOI
Measurement of protein using bicinchoninic acid
TL;DR: This new method maintains the high sensitivity and low protein-to-protein variation associated with the Lowry technique and demonstrates a greater tolerance of the bicinchoninate reagent toward such commonly encountered interferences as nonionic detergents and simple buffer salts.
Book
Methods of Enzymatic Analysis
TL;DR: Methods of enzymatic analysis, Methods of enzymes analysis, the authors, Methods of enzyme analysis, enzymatics, methods of enzymes, and methods of analysis, method of enzymes.
Journal ArticleDOI
A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples
TL;DR: The original Lowry method of protein determination has been modified by the addition of sodium dodecyl sulfate in the alkali reagent and an increase in the amount of copper tartrate reagent to be used with membrane and lipoprotein preparations without prior solubilization or lipid extraction.