Fluorescence detection and quantitation of recombinant proteins containing oligohistidine tag sequences directly in sodium dodecyl sulfate-polyacrylamide gels

Two fluorophore-nitrilotriacetic acid conjugates, Pro-Q Sapphire 365 and Pro-Q Sapphire 488 oligohistidine gel stains, have been developed for the fluorescence detection of fusion proteins containing oligohistidine tags directly in sodium dodecyl sulfate polyacrylamide gels, without the requirement for electroblotting, reporter enzymes or secondary detection reagents. Pro-Q Sapphire 365 oligohistidine gel stain exhibits bright-blue fluorescence (emission maximum = 450 nm) when illuminated with UV-A or UV-B light from a standard ultraviolet transilluminator. Pro-Q Sapphire 488 oligohistidine gel stain exhibits bright-green fluorescence (emission maximum = 515 nm) when illuminated with visible light from a laser-based gel scanner equipped with a 470 nm second-harmonic generation (SHG) or 488 nm argon-ion laser source. Typically, 25-65 ng of oligohistidine-tagged fusion protein in whole cell lysates is detectable using either stain. After documenting the fluorescence signal from the Pro-Q Sapphire dyes, gels may be post-stained with the red-fluorescent SYPRO Ruby protein gel stain in order to reveal the total protein pattern.     

Acylation and Sulfonation of the Enol Form of Phosphorylated Aldehydes

Russian Journal of General Chemistry -     

PHOTOCYTOTOXICITY OF CURCUMIN

Curcumin, bis (4-hydroxy-3-methoxyphenyl)-l,6-diene-3,5-dione, is a yellow-orange dye derived from the rhizome of the plant Curcuma longa. Curcumin has demonstrated phototoxicity to several species of bacteria under aerobic conditions (Dahl, T. A., et al. , 1989, Arch. Microbiol. 151 183), denoting photodynamic inactivation. We have now found that curcumin is also phototoxic to mammalian cells, using a rat basophilic leukemia cell model, and that this phototoxicity again requires the presence of oxygen. The spectral and photochemical properties of curcumin vary with environment, resulting in the potential for multiple or alternate pathways for the exertion of photodynamic effects. For example, curcumin photogenerates singlet oxygen and reduced forms of molecular oxygen under several conditions relevant to cellular environments. In addition, we detected carbon-centered radicals, which may lead to oxidation products (see accompanying paper). Such products may be important reactants in curcumin's phototoxicity since singlet oxygen and reduced oxygen species alone could not explain the biological results, such as the relatively long lifetime (t¹²= 27 s) of the toxicant responsible for decreased cell viability.     

Preparation and reactivity of metal-containing monomers

Russian Chemical Bulletin - The monomer [Cr3O(OCOCH=CH2)6]OH was produced by reaction of acrylic acid with fresh chromium hydroxide. This monomer was investigated by a number of physicochemical...     

Intermediates in the photochemical reaction for the formation of 2,7-dimethylacridine from ditolylamine and bromoform

The reaction of the photochemical synthesis of 2,7-dimethylacridine from di(para-tolyl)amine and bromoform was studied utilizing the method of microsecond impulse photolysis. The proposed new scheme for the formation of acridine assumes the recombination of the primary alkyl radicals at the nitrogen atom of the cation radical and/or the radical of the amine as the main route of the reaction. The data obtained indicate the common character of the nature of some intermediate stages and the structures of the intermediates in the photochemical formation of acridine compounds and triphenylmethane dyes from aromatic amines and halogenomethanes.Institute of Chemical Physics, Chernogolovka, Russian Academy of Sciences, 142432 Chernogolovka. Translated from Izvestiya Akademii Nauk, Seriya Khimicheskaya, No. 9, pp. 2044–2052, September, 1992.     

Intracavity laser polarization spectrometer for biomolecule traces

A polarization variant of an intracavity laser spectrometer based on the competition of beams in an argon ion laser, radiating a number of narrow lines in the 275 to 529 nm spectral range, is proposed and described for the spectral analysis of liquid samples, having broad absorption bands, mainly biomolecules in solutions. By careful optimization of the operating conditions, a procedure has been established that results in a minimal measurable absorbance decreased up to 10(-5) units of optical density. The examined spectrometer provides a wide dynamic range of analysis. A determination of trace contents of amino acids in methanol were carried out.     

Enzymatic polymerizations using surfactant microstructures and the preparation of polymer-ferrite composites

Applied Biochemistry and Biotechnology - Horseradish peroxidase has been used as a biocatalyst to synthesize a polymeric material from alkyl-substituted phenols. The synthesis is carried out in a...     

Collaborative monitoring study of airborne particulate matters among seven Asian countries

Summary Seven Asian countries have been collaborating in collecting airborne particulate matter (APM) in their individual countries and analyzing them by neutron activation analysis as a common analytical tool. APM samples were collected into two fractions of fine and coarse grains (PM2 and PM2-10, respectively). Analytical data were compared from several viewpoints such as particulate sizes, locality of sampling sites (either urban or rural) and geographical location of participating countries. Chemical composition and their monthly variations as well as mass concentrations appear to be highly characteristic for individual sampling sites, suggesting that NAA data are suitable for evaluating the air quality in each site.