A simple enzymic method for the synthesis of [32P]phosphoenolpyruvate.
Ontology highlight
ABSTRACT:
SUBMITTER: Parra F
PROVIDER: S-EPMC1158534 | biostudies-other | 1982 Sep
REPOSITORIES: biostudies-other
Similar Datasets
Project description:A simple, rapid and inexpensive method is described for the enzymic synthesis of [alpha-32P]ATP from [32P]Pi on a preparative scale with an overall yield of 53%. The final product contained all of the detectable radioactivity (less than 99.9%) in the alpha position and has been shown to behave identically with commerically availabe [alpha-32P]ATP during the synthesis of 3':5'-cyclic AMP in the reaction catalysed by adenylate cyclase.
| S-EPMC1164541 | biostudies-other
Project description:Over the past three decades, DNA-encoded library (DEL) technologies have become one of the most relevant strategies for hit-finding. Recent advances in synthetic methodologies for DNA-encoded libraries rendered the increased chemical space available, but it is unknown how every variety of chemistry affects DNA's integrity. Available assays to quantify DNA damage are restricted to electrophoresis, ligation efficiency, and mostly qPCR quantification and sequencing, which may contain predisposition and inconsistency. We developed an external standard method through LC-MS analysis to accurately quantify DNA damage throughout the chemical transformations. An assessment was conducted on on-DNA chemical reactions that are frequently employed in DEL synthesis, and these results were compared to traditional qPCR measurements. Our study provides a simple, practicable, and accurate measurement for DNA degradation during DEL synthesis. Our finding reveals substantial disagreement among the usual DNA-damaging assessment methods, which have been largely neglected so far.
| S-EPMC10733999 | biostudies-literature
Project description:A simple and rapid assay method is described for determining the specific radioactivity of the gamma-phosphate group of 32P-labelled ATP. Labelled ATP is incubated with cyclic AMP, cyclic AMP-dependent protein kinase and histone H2A under conditions leading to maximum phosphorylation of the histone. The specific radioactivity of the gamma-phosphate group of the 32P-labelled ATP is calculated from the total amount of [32P]Pi incorporated into a standard amount of histone. This assay method uses inexpensive commercially available materials, and it yields an accurate specific radioactivity with as little as 0.25 nmol of 32P-labelled ATP.
| S-EPMC1154282 | biostudies-other
Project description:Copper nanoparticles have attracted a wide attention because of their low cost and high specific surface area. At present, the synthesis of copper nanoparticles has the problems of complicated process and environmentally unfriendly materials like hydrazine hydrate and sodium hypophosphite that would pollute water, harm human health and may even cause cancer. In this paper, a simple and low-cost two-step synthesis method was used to prepare highly stable and well-dispersed spherical copper nanoparticles in solution with a particle size of about 34 nm. The prepared spherical copper nanoparticles were kept in solution for one month without precipitation. Using non-toxic l-ascorbic acid as the reducing and secondary coating agent, polyvinylpyrrolidone (PVP) as the primary coating agent, and NaOH as the pH modulator, the metastable intermediate CuCl was prepared. Due to the characteristics of the metastable state, copper nanoparticles were rapidly prepared. Moreover, to improve the dispersibility and antioxidant, the PVP and l-ascorbic acid were used to coat the surface of copper nanoparticles. Finally, the mechanism of the two-step synthesis of copper nanoparticles was discussed. This mechanism mainly relies on the two-step dehydrogenation of l-ascorbic acid to obtain copper nanoparticles.
| S-EPMC10171199 | biostudies-literature
Project description:A new simple and reliable spectrophotometric method is described to determine glyoxylic acid in its synthesis reaction mixture containing oxalic acid, glycolic acid, acetic acid, glyoxal, and ethylene glycol by means of a modified Hopkins-Cole reaction between glyoxylic acid and tryptophan in presence of ferric chloride and concentrated sulphuric acid. The linear range of glyoxylic acid concentration is 0-0.028 M. The limits of detection (LOD) and quantitation (LOQ) are 0.0019 M and 0.00577 M, respectively. The LOD, LOQ, standard deviation, relative standard deviation, and recovery ratio of the proposed method are comparable with a selected HPLC reference method. Both methods displayed same precision and credibility. Reaction stoichiometry between tryptophan and glyoxylic acid is assumed to be 2 : 3. Reaction mechanism has been postulated based on identified molar ratios of reactants. Glyoxal gave a negative test with tryptophan although it is a dialdehyde.
| S-EPMC7155763 | biostudies-literature
Project description:CRISPR-Cas9 sgRNA libraries have transformed functional genetic screening and have enabled several innovative methods that rely on simultaneously targeting numerous genetic loci. Such libraries could be used in a vast number of biological systems and in the development of new technologies, but library generation is hindered by the cost, time, and sequence data required for sgRNA library synthesis. Here, we describe a rapid enzymatic method for generating robust, variant-matched libraries from any source of cDNA in under 3 h. This method, which we have named SLALOM, utilizes a custom sgRNA scaffold sequence and a novel method for detaching oligonucleotides from solid supports by a strand displacing polymerase. With this method, we constructed libraries targeting the E. coli genome and the transcriptome of developing zebrafish hearts, demonstrating its ability to expand the reach of CRISPR technology and facilitate methods requiring custom libraries.
| S-EPMC8682767 | biostudies-literature
Project description:A novel method to separate [3H]Ins(1,4,5)P3 and [3H]Ins(1,3,4)P3 in tissue extracts is described. It is based on the selective metabolism of Ins(1,3,4)P3 by a crude cerebral supernatant in a Mg2+-free buffer followed by separation of [3H]inositol trisphosphates using conventional anion-exchange chromatography. Evaluation of the assay was performed using [3H]Ins(1,3,4)P3 standards and tissue extracts containing different proportions of [3H]Ins(1,4,5)P3 and [3H]Ins(1,3,4)P3. Parallel h.p.l.c. separations of extracts established the selective and complete metabolism of [3H]Ins(1,3,4)P3 under the above conditions and demonstrated that the enzymic method provides an accurate estimate of the trisphosphate isomers in rat cerebral cortex, parotid gland and bovine tracheal smooth muscle.
| S-EPMC1138658 | biostudies-other
Project description:In this paper, we report a simple two-step approach for the synthesis of large graphene oxide (GO) sheets with lateral dimensions of ≈10 μm or greater. The first step is a pretreatment step involving electrochemical exfoliation of graphite electrode to produce graphene in a mixture of H2SO4 and H3PO4. The second step is the oxidation step, where oxidation of exfoliated graphene sheets was performed using KMnO4 as the oxidizing agent. The oxidation was carried out for different times ranging from 1 to 12 h at ∼60 °C. Prepared GO batches were characterized using a number of spectroscopy and microscopy techniques such as X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), and UV-visible spectroscopy. Raman and thermogravimetric analysis techniques were used to study the degree of oxidation in the as-synthesized GO batches. The UV-visible absorption spectrum showed an intense peak at 230 nm and an adjacent band at 300 nm corresponding to π-π* and n-π* transitions in all samples. Normalized FTIR plots were used to calculate the relative percentages of oxygen-containing functional groups, which were found to be maximum in GO (6 h). Boehm titration was used to quantify the functional groups present on the GO surface. Overall GO sheets obtained after 6 h of oxidation, GO (6 h), were found to be the best. XRD pattern of GO (6 h) revealed a characteristic peak at 2θ = 8.88°, with the corresponding interplanar spacing between the layers being 0.995 nm, which is among the best with respect to the previous methods reported in the literature. Raman spectroscopy showed that the degree of defect (I D/I G) area ratio for GO (6 h) was 1.24, which is higher than that obtained for GO (1.18) prepared by widely used Marcano's approach.
| S-EPMC6645570 | biostudies-literature
Project description:Cyclodextrin (β-CD)-functionalized reduced graphene oxide was successfully synthesized by a simple hydrothermal method, followed by conjugating with polyethylene glycol (PEG) and folic acid (FA). Microscopic and spectroscopic techniques were used to characterize the nanocomposites. Photothermal experiments showed that β-CD-functionalized reduced graphene oxide exhibited higher photothermal conversion efficiency in the near infrared region than reduced graphene oxide functionalized with other molecules under the same conditions. Cytotoxicity experiments indicated that rGO@CD@PEG@FA possessed good biocompatibility even at high concentration. When doxorubicin (DOX) was loaded on the rGO@CD@PEG@FA nanocomposite, it showed the stimulative effect of heat, pH response, and sustained drug release. Cytotoxicity experiments also confirmed the targeted effect and high efficiency of the combined therapy. The findings of the present study provide an ideal drug delivery system for malignant cancer therapy due to the advanced synergistic chemo-photothermal targeted therapy and good drug release properties.
| S-EPMC9089399 | biostudies-literature
Project description:A rapid practical process has been developed for synthesis of 2,4,5-trisubstituted-imidazoles in excellent yields up to 95% from readily available starting materials. In this CuI catalyzed synthesis, trisubstituted imidazoles were afforded in short reaction times, wherein the substrate scope is well explored with benzoin as well as benzil reacting with different aldehydes in the presence of ammonium acetate as the nitrogen source.
| S-EPMC9034145 | biostudies-literature