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
Comparison of the physicochemical properties of a biosimilar filgrastim with those of reference filgrastim.
Ana Skrlin,I. Radic,Marko Vuletić,D. Schwinke,D. Runac,T. Kusalic,I. Paskvan,M. Krsic,M. Bratos,S. Marinc +9 more
TL;DR: Data show that the physicochemical profile of Hospira filgrastim is similar to that of Amgen filgrastsim, which are shown to have similar physicochemical properties, molecular characteristics, purity and biological activity.
Journal ArticleDOI
A new strategy for aromatic ring alkylation in cylindrocyclophane biosynthesis.
TL;DR: Cylindrocyclophane biosynthesis in Cylindrospermum licheniforme ATCC 29412 involves chlorination of an unactivated carbon center by a novel halogenase, followed by a previously uncharacterized enzymatic dimerization reaction featuring sequential, stereospecific alkylations of resorcinol aromatic rings.
Journal ArticleDOI
Functional expression of a thermophilic glucuronoyl esterase from Sporotrichum thermophile : identification of the nucleophilic serine
TL;DR: A glucuronoyl ester enzyme from the thermophilic fungus Sporotrichum thermophile, belonging to the carbohydrate esterase family 15 (CE-15), was functionally expressed in the methylotrophic yeast Pichia pastoris, rendering StGE2 a strong candidate for future biotechnological applications that require robust biocatalysts.
Journal ArticleDOI
Development of a fast, reproducible and effective method for the extraction and quantification of proteins of micro-algae
E.A. Meijer,R.H. Wijffels +1 more
TL;DR: The extraction method was the most critical factor factor for determination of proteins in micro-algae and needs to be optimized for each algal species.
Journal ArticleDOI
The use of inorganic elemental standards in the quantification of proteins and biomolecular compounds by inductively coupled plasma spectrometry
TL;DR: In this article, an inductively coupled plasma atomic emission spectrometer (ICP-AES) and an ICP-time-of-flight mass spectrometers were investigated in terms of their generic behaviour, i.e. if their responses can be considered as structure independent or not in the simple matrices prevailing during a liquid chromatographic (LC) run.
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.