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Showing papers in "Analytical Biochemistry in 1987"


Journal ArticleDOI
TL;DR: A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described, providing a pure preparation of undegraded RNA in high yield and can be completed within 4 h.

65,881 citations


PatentDOI
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.

20,907 citations


Journal ArticleDOI
TL;DR: A discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) system for the separation of proteins in the range from 1 to 100 kDa is described, and the omission of glycine and urea prevents disturbances which might occur in the course of subsequent amino acid sequencing.

11,290 citations


Journal ArticleDOI
TL;DR: The universal method to detect and determine siderophores was developed by using their high affinity for iron(III) and was successfully used to screen mutants in the iron uptake system of two Rhizobium meliloti strains, DM5 and 1021.

5,499 citations


Journal ArticleDOI
TL;DR: The effects of experimental variables on assays based upon the use of either ferricytochrome c or nitro blue tetrazolium are investigated to help investigators to avoid the numerous potential pitfalls which necessarily surround these assay methods.

2,342 citations


Journal ArticleDOI
TL;DR: It is suggested that the deoxyribose assay is a simple and cheap alternative to pulse radiolysis for determination of rate constants for reaction of most biological molecules with hydroxyl radicals.

2,296 citations


Journal ArticleDOI
TL;DR: A fast and efficient method for the isolation of RNA from plant tissues is described, of particular use for isolating RNA from tissues with a high polysaccharide and nuclease content such as wounded potato tubers.

1,812 citations


Journal ArticleDOI
TL;DR: Two procedures for analyzing overlapping optical spectra of mixtures of pyridine hemochromes are described, and extinction coefficients of pyrsidine hemochemes are provided for use with these methods, and these two procedures are generally applicable to analyses of absolute or difference specta of mixture of components obeying Beer's law.

851 citations


Journal ArticleDOI
TL;DR: A procedure, based on the complex formation of malachite green with phosphomolybdate under acidic conditions, to measure inorganic orthophosphate in the nanomolar range is described, with advantages of simplicity, stability of the reagents, and high sensitivity.

713 citations


Journal ArticleDOI
TL;DR: Although the variable selection method improves the analysis of circular dichroism data truncated at 190 nm, data measured to 178 nm gives superior results, it is shown that improving the fit to the measured CD beyond the accuracy of the data can result in poorer analyses.

694 citations


Journal ArticleDOI
TL;DR: The dye Nile red detects the exposure or formation of new hydrophobic surfaces induced by ligand binding to calmodulin, oligomerization of melittin, or unfolding of ovalbumin during early thermal denaturation.

Journal ArticleDOI
TL;DR: The procedure permits the application of larger sample volumes containing more protein than other isoelectric focusing procedures, and provides the additional advantages of slab gels over tube gels for analytical purposes, making it possible to complete a two-dimensional gel in 1 day.

Journal ArticleDOI
TL;DR: It is anticipated that CuCl2 will be useful in the rapid analysis of proteins by polyacrylamide gel electrophoresis and in the preparation of purified polypeptides by elution from gel slices.

Journal ArticleDOI
TL;DR: A new method by combining an HPLC system with an ultrasensitive analytical system based on the detection of chemiluminescence emitted by isoluminol in the presence of hydroperoxide and microperoxidase to detect lipid hydroperoxides at the picomole level is developed.

Journal ArticleDOI
TL;DR: This investigation demonstrates that Stractan density gradient centrifugation is an efficient, sensitive, and reproducible method for isolating pure populations of hepatic nonparenchymal cells.

Journal ArticleDOI
TL;DR: Normal ranges for total homocysteine, total cystine, and methionine have been determined in human and rat serum and in human urine.

Journal ArticleDOI
TL;DR: A simple, rapid, and highly sensitive high-performance liquid chromatographic method is described for the determination of N-acetyl- and N-glycolylneuraminic acids in human and animal sera, glycoproteins, and glycolipids.

Journal ArticleDOI
TL;DR: The photometric assay of reducing sugars with 2,2'-bicinchoninate was improved and its conditions were optimized, and the molar extinction coefficients produced with the 12 most common monosaccharides occurring in carbohydrates and conjugated glycocompounds are listed.

Journal ArticleDOI
TL;DR: Assays for the in vitro metabolism of (+)- and (-)-bufuralol, debrisoquine, and dextromethorphan in human liver microsomes and reconstituted purified cytochrome P-450 isozymes indicate the involvement of the same enzyme in the metabolism of all three substrates investigated.

Journal ArticleDOI
TL;DR: Inclusion of guanidinium chloride in the assay resulted in a stable dye-proteoglycan interaction, but eliminated the interference of other anionic macromolecules such as DNA.

Journal ArticleDOI
TL;DR: The efficiency of genetic transformation of Saccharomyces cerevisiae sp Heroplasts has been increased 10- to 100-fold over previously published procedures, indicating that fusion is not required for the uptake of DNA by yeast spheroplasts.

Journal ArticleDOI
TL;DR: As more investigators try the matrices and matrix components described, as well as new components and combinations of them, it is anticipated that improvement in the culture of many cells can be expected.

Journal ArticleDOI
TL;DR: Intact and readily digestible DNA was prepared from a range of murine tissues and cell lines which had been dispersed in GHCl and ethanol precipitated, and then rinsed in 70% ethanol and resuspended in 10 mM Tris-HCl, 1 mM EDTA, pH 7.5.

Journal ArticleDOI
TL;DR: The method described here used a specially designed mobile phase, gave greater stability and a noiseless baseline, and increased substantially the sensitivity and precision.

Journal ArticleDOI
TL;DR: A simple and fast ion pair reversed-phase high-performance liquid chromatographic method has been developed for the simultaneous determination of ATP, ADP, AMP, GTP, GDP, IMP, NADP+, NADPH+, NAD+, NADH,ADP-ribose, inosine, adenosine, hypoxanthine, and xanthine.

Journal ArticleDOI
TL;DR: The results indicate that blood for DNA extraction may be mailed in an ambient temperature container and, in many cases, sent by first-class mail rather than by overnight delivery services.

Journal ArticleDOI
TL;DR: Endoglucanase and cellobiohydrolase components of thermophilic cellulases can be detected in situ after gel electrophoresis in the presence of sodium dodecyl sulfate by incorporating a mixed linkage beta-glucan (barley beta- glucan, lichenan) in the separation gel.

Journal ArticleDOI
TL;DR: This work has developed a method for determining oxidized and reduced NAD and NADP in human erythrocytes using a single extract that eliminates the need for reducing agents and thus allows the use of a spectrophotometric cycling assay.

Journal ArticleDOI
Pål Nyrén1
TL;DR: A simple and rapid method has been developed for continuous monitoring of DNA synthesis in vitro, and the effect of an inhibitor, adriamycin, on the polymerase activity was studied.

Journal ArticleDOI
TL;DR: A paired-ion, reversed-phase, high-performance liquid chromatography procedure using electrochemical detection and internal standard quantitation based on isoascorbic acid (IA), which overcomes a major problem associated with AA instability and eliminates the necessity of assaying samples immediately after they are prepared for analysis.