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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Book ChapterDOI
Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent
TL;DR: Aggregate analysis of this type is an important supplement to and often more informative than reems of data difficult to summarize from various techniques, such as high-performance liquid chromatography (HPLC) that separate a large number of individual compounds.
Journal ArticleDOI
Nano-C60 cytotoxicity is due to lipid peroxidation.
Christie M. Sayes,Andre M. Gobin,Kevin D. Ausman,Joe S. Mendez,Jennifer L. West,Vicki L. Colvin +5 more
TL;DR: With the addition of an antioxidant, L-ascorbic acid, the oxidative damage and resultant toxicity of nano-C60 was completely prevented and damage to cell membranes was observed both with chemical assays, and confirmed physically by visualizing membrane permeability with high molecular weight dyes.
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Nitrate reductase-mediated synthesis of silver nanoparticles from AgNO3
S. Anil Kumar,Majid Kazemian Abyaneh,Suresh W. Gosavi,Sulabha K. Kulkarni,Renu Pasricha,Absar Ahmad,M. I. Khan +6 more
TL;DR: These studies will help in designing a rational enzymatic strategy for the synthesis of nanomaterials of different chemical composition, shapes and sizes as well as their separation.
Journal ArticleDOI
A 'proteomic ruler' for protein copy number and concentration estimation without spike-in standards
TL;DR: This work shows that the MS signal of histones can be used as a “proteomic ruler” because it is proportional to the amount of DNA in the sample, which in turn depends on the number of cells, and adds an absolute scale to the MS readout and allows estimation of the copy numbers of individual proteins per cell.
Journal ArticleDOI
A Tomato Peroxidase Involved in the Synthesis of Lignin and Suberin
Mónica Quiroga,Consuelo Guerrero,Miguel A. Botella,Araceli Barceló,Iraida Amaya,María I. Medina,Francisco J. Alonso,Silvia Milrad de Forchetti,Horacio A. Tigier,Victoriano Valpuesta +9 more
TL;DR: It is proposed that the TPX1 product is involved in the synthesis of lignin and suberin, a tomato peroxidase gene the authors have previously isolated.
References
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Journal ArticleDOI
Lowry protein assay using an automatic microtiter plate spectrophotometer.
TL;DR: The method of protein determination reported by Lowry et al. has been adapted for use with 96-well microtiter plates and an automatic microplate spectrophotometer and offers advantages over previously reported methods in that it is more rapid and uses a smaller sample volume.
Journal ArticleDOI
Simple, rapid, turbidometric determination of inorganic sulfate and/or protein.
S.G. Jackson,E.L. McCandless +1 more
TL;DR: A simple, reproducible, turbidometric assay, adaptable to protein and inorganic sulfate analysis, which develops over a relatively short time and is stable for some time (over an hour for sulfateAnalysis).
Journal ArticleDOI
A linear standard curve for the Folin Lowry determination of protein.
Journal ArticleDOI
Measurement of protein in cell suspensions using the Commassie brilliant blue dye-binding assay
G.O. Gogstad,M. B. Krutnes +1 more
TL;DR: It is shown that the addition of small amounts of Triton X-100 or NaOH to the cell suspensions prior to addition of the dye reagent corrected the discrepancy in direct measurement of protein in cell suspensions using the Coomassie brilliant blue dye-binding assay.
Journal ArticleDOI
Adaptation of the dye-binding protein assay to microtiter plates.
TL;DR: The dye-binding protein assay has been adapted for use with microtiter plates and a plate reader, and the response of nitrate reductase in the assay is about 70% of that of the standard protein, bovine serum albumin.