Showing papers in "Analytical Biochemistry in 1989"

TL;DR: In this article, a method for calculating accurate molar extinction coefficients for proteins at 280 nm, simply from knowledge of the amino acid composition, was presented, and the method was calibrated against 18 "normal" globular proteins.

TL;DR: A new titration calorimeter is described and results are presented for the binding of cytidine 2'-monophosphate (2'CMP) to the active site of ribonuclease A.

TL;DR: Fluorescence intensity in the presence of a constant amount of amyloid fibrils reached a plateau with increase in the thioflavine T concentration, and the fluorescence was considerably diminished when structure of the amyloids fibril was disrupted by guanidine-HCl treatment.

TL;DR: Passees en revue les differentes endoglycosidases d'origine bacterienne, animale ou humaine permettant de par leur specificite d'hydrolyse, de differencier les residus glycosyles portes par les glycoproteines et donc de caracteriser ces dernieres.

TL;DR: The modifications described here permit quick, efficient removal of many interfering substances that are commonly utilized during protein purification.

TL;DR: The inclusion of BCS in the NBT assay system was shown to make the accurate measurement of SOD activity in tissues with interferences possible, and the most effective compounds were metal chelators that were also electron transport chain inhibitors.

TL;DR: The method for cell number measurement in monolayer cultures by crystal violet staining published recently was modified and significantly improved and it is shown that the sensitivity of the assay can be varied and easily controlled by choosing the appropriate pH during the staining procedure.

TL;DR: Reverse-phase partition chromatography on a column of Capcell Pak C18 with uv or electrochemical detection allowed rapid analysis of aldoses and N-acetylhexosamines with the detection limit of 1 pmol or 100 fmol, respectively, and proved especially useful for analysis of component monosaccharides of glycorproteins.

TL;DR: Commercial Streptomyces griseus and Serratia marcescens chitinases and purified wheat germ W1A and hen egg white lysozymes were subjected to polyacrylamide gel electrophoresis under native conditions at pH 4.3 and molecular weights of chitinolytic enzymes could be directly estimated.

TL;DR: Immobilon, a membrane of polyvinylidene difluoride to which gel-fractionated proteins can be transferred electrophoretically, was found to be an excellent matrix for the analysis of the phosphoamino acid content of phosphoproteins.

TL;DR: When microtubule-associated protein 2 was included in the gel, the sensitivity of the detection of calmodulin-dependent protein kinase II in the Gel was more than one order of magnitude higher than that in thegel containing no protein substrate.

TL;DR: A procedure for covalent immobilization of functional proteins on silica substrates was developed using thiol-terminal silanes and heterobifunctional cross-linkers and is straightforward, easy to perform, and adaptable for modifying mass quantities of biosensor components.

TL;DR: This method of site-specific mutagenesis can be applied to any linear double-stranded DNA large enough for primer annealing and obviates specialized cloning vectors, DNA constructs, and selection techniques and has the advantage over a recently published PCR technique.

TL;DR: The purpose of this review is to examine the merits of the major groups of currently available viability assays as they apply to mammalian cell systems and to show that while some assays may be clearly superior in measuring a final endpoint, technical and practical considerations may limit their usefulness.

TL;DR: A procedure is presented for accurate determination by ion exchange chromatography and postcolumn derivatization of all amino acids from acid hydrolysis of a protein, including the Cys-derivative.

TL;DR: A method to quantify oxidatively modified proteins through reduction of these carbonyl groups with tritiated borohydride is described, which provided excellent sensitivity and was directly proportional to the amount of protein.

TL;DR: The use of 96-well microtiter plates and a programmable microplate reader to measure glutathione reductase in an assay based on reduction of 5,5'-dithiobis(2-nitrobenzoic acid) by GSH generated from an excess of GSSG is described.

TL;DR: DNA was assayed in a homogeneous format using DNA probes containing hybridization-sensitive labels that were able to detect unlabeled target DNA by competitive hybridization producing fluorescence signals which increased with increasing target DNA concentration.

TL;DR: A simple, rapid, and highly sensitive fluorometric high-performance liquid chromatographic method for the determination of N- acetylneuraminic acid and its mono-O-acetyl derivatives in human and rat sera is described.

TL;DR: A frequency distribution of amino acids bracketing approximately 400 transmembrane peptide sequences showed Cys to be the least frequently occurring amino acid at this putative interfacial membrane region, indicating Cys at the interface of cells may stabilize protein-lipid interactions.

TL;DR: Inclusion of 10 mM 2-mercaptoethanol in the upper reservoir buffer or blocking free sulfhydryl groups with N-ethylmaleimide prevented intermolecular disulfide bond formation during electrophoresis and improved efficiency of protein transfer to nitrocellulose.

TL;DR: On-column uv monitoring and quantification by the relative peak area method allowed reproducible determination of these saccharides at least in the concentration range of 10-100 mM in reaction solutions, and this method was applied to the determination of the monosaccharide compositions of various carbohydrate materials.

TL;DR: Initial results combining rapid separations by capillary zone electrophoresis with on-line mass spectrometric detection via the electrospray ionization source are demonstrated, and the potential for extension of these methods to molecular weights on the order of 10(6) is discussed.

TL;DR: Evidence is provided to show that in the conventional phenol-sulfuric acid reaction procedure, phenol underwent sulfonation in situ and the phenolsulfonic acid formed decreased the color intensity for hydroxymethyl furfural (HMF), furF, and many hexoses and pentoses tested, and a modified method to overcome this problem was described.

TL;DR: An excellent parallelism between hydrolysis determined by a classical extraction method and that followed by direct and continuous spectrofluorometric recording of the monomer emission of pyrene is obtained.

TL;DR: A greater than 40-fold difference in rates of chlorpyrifosoxon hydrolysis observed between rat (low activity) and rabbit sera (high activity) correlated well with the reported large differences in LD50 values for chlorp Pyrifos in these two animals, consistent with an important role of serum paraoxonase in detoxification of organophosphorus pesticides in vivo.

TL;DR: This modified assay for deoxyribonucleoside triphosphates has been improved by using synthetic oligonucleotides of a carefully defined sequence as template primers for DNA polymerase to obtain the lowest background reaction and highest sensitivity.

TL;DR: DNA prepared by this new method is an excellent substrate for restriction endonuclease digestion and Southern hybridization analysis and sodium perchlorate and chloroform instead of phenol with a significant time savings realized as well as fewer hazards to the technician.

TL;DR: The physical, optical, and anion quenching properties for the series of ester compounds were determined to establish a set of structure-activity correlates.

TL;DR: The effect of noncharged neutral amino acids on mobility was investigated and shown to be present, but only as a minor perturbation on the effects of size and charge.