Book ChapterDOI
Quantitation of protein.
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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
Cocaine Detoxification by Human Plasma Butyrylcholinesterase
Thomas J. Lynch,Carol E. Mattes,Anu Singh,Roy M. Bradley,Roscoe O. Brady,Kenneth L. Dretchen +5 more
TL;DR: Investigation of the ability of human plasma butyrylcholinesterase (BChE) to detoxify cocaine in vivo suggested that BChE would provide a safe and highly efficacious treatment for cocaine intoxication.
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TNFα induces expression of the chemoattractant cytokine RANTES in cultured mouse mesangial cells
Gunter Wolf,Stefan Aberle,Friedrich Thaiss,Peter J. Nelson,Alan M. Krensky,Eric G. Neilson,Rolf A.K. Stahl +6 more
TL;DR: The results indicate that mesangial cells produce the small cytokine RANTES, which, in concert with other chemoattractants, may play a role in the glomerular recruitment of inflammatory cells like macrophages/monocytes.
Journal ArticleDOI
Activation of the phosphosignaling protein CheY. II. Analysis of activated mutants by 19F NMR and protein engineering.
TL;DR: Results are consistent with a two-step model of activated CheY docking to the flagellar switch, and the perturbation of the Lys109 region observed for activating mutations is not detected in the phosphorylated protein.
Journal ArticleDOI
DNAJB6 is a peptide-binding chaperone which can suppress amyloid fibrillation of polyglutamine peptides at substoichiometric molar ratios
Cecilia Månsson,Vaishali Kakkar,Elodie Monsellier,Yannick Sourigues,Johan Härmark,Harm H. Kampinga,Ronald Melki,Cecilia Emanuelsson +7 more
TL;DR: The data obtained in cells and in vitro support the view that DNAJB6 is a peptide-binding chaperone that can interact with polyQ peptides that are incompletely degraded by and released from the proteasome.
Journal ArticleDOI
The maize brittle 1 gene encodes amyloplast membrane polypeptides.
Thomas D. Sullivan,Yasuko Kaneko +1 more
TL;DR: During endosperm fractionation by differential centrifugation and membrane sedimentation in sucrose gradients, the Bt1 proteins co-purified with the carotenoid-containing plastid membranes and were localized to amyloplasts by electron-microscopic immunocytochemistry; most of the signal was detected at the plastids periphery.
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.