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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ATPase kinetics for wild‐type Saccharomyces cerevisiae F1‐ATPase and F1‐ATPase with the β‐subunit Thr197→Ser mutation
TL;DR: The results indicate that the effects of azide, oxyanions, and the Thr197-->Ser mutation on the biochemistry of F1-ATPase are limited primarily to multisite conditions.
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Immunization with a recombinant fusion protein protects mice against Helicobacter pylori infection.
Amir Ghasemi,Nazanin Mohammad,Josef Mautner,Mehrnaz Taghipour Karsabet,Jafar Amani,Abolfazl Ardjmand,Zarichehr Vakili +6 more
TL;DR: RFUVL immunized mice exhibited a reduction of about three orders of magnitude in their stomach bacterial loads, and might be considered as a promising vaccine candidate for the control of H. pylori infection.
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A calcium-activated nucleotidase secreted from Ostertagia ostertagi 4th-stage larvae is a member of the novel salivary apyrases present in blood-feeding arthropods.
TL;DR: A gene (Oos-apy-1) encoding a protein that structurally conforms to the Ca2+-activated apyrase from the bed bug, Cimex lectularius, is identified by immunologically screening an Ostertagia L4 cDNA expression library.
Journal ArticleDOI
Biochemical properties of 5'-nucleotidase from mouse skeletal muscle.
Alejandro Martínez-Martínez,César Flores-Flores,Francisco J. Campoy,Encarnación Muñoz-Delgado,Carlo Fini,Cecilio J. Vidal +5 more
TL;DR: Ecto-5'-nucleotidase from mouse muscle has been purified after extraction with detergent followed by chromatography on concanavalin A- and AMP-Sepharose and revealed that enzyme forms in F-I or F-II were amphiphilic dimers with linked phosphatidylinositol residues, whilst most of NT forms in UF were hydrophilic dimer.
Journal ArticleDOI
Telomerase activity is upregulated in the fat bodies of pre-diapause bumblebee queens (Bombus terrestris)
Justina Koubová,Tomáš Jehlík,Dalibor Kodrík,Michala Sábová,Peter Šima,Hana Sehadova,Radka Závodská,Radmila Čapková Frydrychová,Radmila Čapková Frydrychová +8 more
TL;DR: In somatic tissues, telomerase activity was upregulated only in the fat bodies of pre-diapause queens, and this upregulation was linked to heightened DNA synthesis, and the lifespan differences between B. terrestris workers and queens are related to the long diapause period of the queen.
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.