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Yue Liu, Jia Hu, Yan Li, Yun-Tao Shang, Jia-Qi Wang, Ye Zhang, Zhong-Liang Wang
In this work, a CE method was developed to separate five anthraquinones: aloe-emodin, rhein, emodin, chrysophanol and physcion. The CE method used a nano-sized metal organic framework MIL-101 (nMIL-101) as pseudostationary phase (PSP) and sorbent for dispersed particle extraction (DPE). The nMIL-101 was synthesized by microwave technique and was characterized by UV-vis, TEM, Zeta potential, X-ray diffraction spectrometry and micropore physisorption. In this method, anthraquinones were adsorbed by nMIL-101 of a fast kinetics within 10 min and then separated by CE...
July 18, 2017: Electrophoresis
Jun-Hua Gong, Jian-Ping Gong, Kai-Wen Zheng
The ability to determine that successful transfer and equal loading occur prior to using primary antibodies is important. And total protein staining is commonly used to check transfer efficiency and normalization, which play a crucial role in western blotting. Ponceau S and coomassie blue are commonly used, but, there are disadvantages reported in recent years. Therefore, we are interested in finding another method, which is cheap, easy and fast. As we know, protein binding region of PVDF membrane is still hydrophilic when carbinol volatilizes, however, the non-protein binding region of PVDF membrane became hydrophobic again...
July 18, 2017: Electrophoresis
Yuhong Mao, Urh Černigoj, Viktor Zalokar, Aleš Štrancar, Ulrich Kulozik
Tryptic hydrolysis of β-Lactoglobulin (β-Lg) is attracting more and more attention due to the reduced allergenicity and the functionality of resulting hydrolysates. To produce hydrolysates in an economically viable way, immobilized trypsin reactors (IMTRs), based on polymethacrylate monolith with pore size 2.1 μm (N1) and 6 μm (N2), were developed and used in a flow-through system. IMTRs were characterized in terms of permeability and enzymatic activity during extensive usage. N1 showed twice the activity compared with N2, correlating well with its almost two times higher amount of immobilized trypsin...
July 17, 2017: Electrophoresis
Gabor Járvás, Tamas Varga, Marton G Szigeti, Laszlo Hajba, Peter Fürjes, Istvan Rajta, Andras Guttman
As a continuation of our previously published work, this paper presents a detailed evaluation of a microfabricated cell capture device utilizing a doubly tilted micropillar array. The device was fabricated using a novel hybrid technology based on the combination of proton beam writing and conventional lithography techniques. Tilted pillars offer unique flow characteristics and support enhanced fluidic interaction for improved immuno-affinity based cell capture. The performance of the microdevice was evaluated by an image sequence analysis based in-house developed single cell tracking system...
July 17, 2017: Electrophoresis
Zhao Chen, Aili Xu, Xiaoli Bi, Wenhui Luo, Ji Li, Guorong Fan, Dongmei Sun
In the present study, a monolithic capillary column with higher permeability was developed for the in-vivo discrimination of four coumarin analogs (bergapten, 2'-acetylangelicin, imperatorin and osthole) that typically require long separation times in HPLC. Instead of conventional methacrylate ester monolith (containing 19.5% porogen) with insufficient permeability (K = 1.52-1.66 × 10(-14) M(2) ) for plasma sample, the proposed column (20.5% porogen) had better permeability (around 3.80 × 10(-14) M(2) ) while properties such as pore distribution, stability and resolution changed slightly...
July 17, 2017: Electrophoresis
Vanessa Martinez, Deepthi Nori, Pero Dimsoski, Bruce McCord
A pressure based protocol for differential extraction has been optimized to provide a rapid and selective alternative to conventional differential extraction techniques with the advantage of an increased recovery of genomic DNA. The protocol involves treating cotton swabs containing mixtures of sperm and vaginal epithelial cells with 2 sets of pressure treatment using a Barocycler(®) NEP 2320 in alkaline conditions. This first step quantitatively and selectively removes female epithelial cells. The cotton swab is removed and further treated with alkali at 95°C for the removal of sperm cell DNA...
July 17, 2017: Electrophoresis
Alessia Ferrarini, Laura Righetti, Ma Paz Martínez, Mariano Fernández, Annalaura Mastrangelo, Juan P Horcajada, Antoni Betbesé, Andrés Esteban, Jordi Ordóñez, Joaquín de Gea, Jesús Ruiz Cabello, Federica Pellati, José A Lorente, Nicolás Nin, Francisco J Rupérez
Acute Respiratory Distress Syndrome (ARDS) is a serious complication of influenza A (H1N1) virus infection. Its pathogenesis is unknown, and biomarkers are lacking. Untargeted metabolomics allows the analysis of the whole metabolome in a biological compartment, identifying patterns associated with specific conditions. We hypothesized that LC-MS could help identify discriminant metabolites able to define the metabolic alterations occurring in patients with influenza A (H1N1) virus infection that developed ARDS...
July 17, 2017: Electrophoresis
Agnieszka Rogowska, Katarzyna Rafińska, Paweł Pomastowski, Justyna Walczak, Viorica Railean-Plugaru, Magdalena Buszewska-Forajta, Bogusław Buszewski
The resistance of pathogenic bacteria to antibiotics has become a serious problem. The emphasis is placed on the development of new, effective antimicrobial strategies. One of them is the use of AgNPs in association with antibiotic drugs. The aim of this study was to obtain silver nanoparticles functionalized with ampicillin and to investigate the mechanism of binding antibiotics to nanoparticle using high-performance liquid chromatography approach. To confirm the occurrence of silver nanoparticles functionalization, FTIR, MALDI-TOF MS, and DLS analysis and zeta potential measurements were performed...
July 13, 2017: Electrophoresis
Kimberly Schwartz, Dmitry Bochkariov
Western blotting is a ubiquitous tool used in protein and molecular biology research, providing information about the presence, size, relative abundance and state of a protein in a mixture. First, the proteins in a sample are separated by size using SDS-PAGE then transferred onto a membrane for detection with a set of primary and secondary antibodies. High-quality Western data requires high signal-to-noise ratios, which depend upon reduction of nonspecific antibody interactions. Blocking is a critical step in the Western blot method as it prevents the antibodies from binding nonspecifically to the membrane and irrelevant proteins...
July 13, 2017: Electrophoresis
Le Si-Hung, Tim J Causon, Stephan Hann
Reversed-phase LC combined with high-resolution mass spectrometry (HRMS) is one of the most popular methods for cellular metabolomics studies. Due to the difficulties in analysing a wide range of polarities encountered in the metabolome, 100%-wettable reversed-phase materials are frequently used to maximise metabolome coverage within a single analysis. Packed with silica-based sub-3 μm diameter particles, these columns allow high separation efficiency and offer a reasonable compromise for metabolome coverage within a single analysis...
July 10, 2017: Electrophoresis
Nadar Khan, Satoru Yamaguchi, Tomoyuki Katsube-Tanaka
Proteolytic cleavage or partial degradation of proteins is one of the important post-translational modifications for various biological processes, but it is difficult to analyze. Previously, we demonstrated that some subunits of the major rice (Oryza sativa L.) seed storage protein glutelin are partially degraded to produce newly identified polypeptides X1-X5 in mutants in which another major seed storage protein globulin is absent. In this study, the new polypeptides X3 and X4/X5 were immunologically confirmed to be derived from GluA3 and GluA1/GluA2 subunits, respectively...
July 6, 2017: Electrophoresis
Jianhao Wang, Jie Fan, Li Liu, Shumin Ding, Xiaoqian Liu, Jianpeng Wang, Liqian Gao, Souvik Chattopadhaya, Peng Miao, Jiang Xia, Lin Qiu, Pengju Jiang
Herein, a novel assay has been developed for monitoring PreScission protease (His-PSP) mediated enzyme cleavage of ATTO 590 labeled peptide substrate (ATTO-LEV). This novel method is based on combining the use of capillary electrophoresis and fluorescence detection (CE-FL) to dynamically monitor the enzyme cleavage activity. A multivalent peptide substrate was first constructed by immobilizing His-tagged ATTO 590 labeled peptide substrate (ATTO-LEVH6) onto the surface of CdSe/ZnS quantum dots (QDs). Once successfully immobilized, the novel multivalent peptide substrate resulted in the Förster resonance energy transfer (FRET) from QDs to ATTO 590...
July 6, 2017: Electrophoresis
LÃcia F A Amorim, Rita Gaspar, Patrà Cia Pereira, Urh Černigoj, Fani Sousa, J A Queiroz, Ãngela Sousa
Affinity chromatography based on amino acids as interacting ligands was already indicated as an alternative compared to ion exchange or hydrophobic interaction for plasmid DNA purification. Understanding the recognition mechanisms occurring between histidine-based ligands and nucleic acids enables more efficient purification of a DNA vaccine, as the binding and elution conditions can be adjusted in order to enhance the purification performance. Decreasing pH to slightly acidic conditions increases the positive charge of histidine ligand, what influences the type of interaction between chromatographic support and analytes...
July 6, 2017: Electrophoresis
Shohei Tazawa, Kazuhiro Kobayashi, Takanori Oyoshi, Masamichi Yamanaka
Pulsed-field gel electrophoresis is a frequent technique used to separate exceptionally large DNA fragments. In a typical continuous field electrophoresis, it is challenging to separate DNA fragments larger than 20 kbp because they migrate at a comparable rate. To overcome this challenge, it is necessary to develop a novel matrix for the electrophoresis. Here, we de-scribe the electrophoresis of large DNA fragments up to 166 kbp using a supramolecular gel matrix and a typical continuous field electrophoresis system...
July 6, 2017: Electrophoresis
Miguel Ángel González-Curbelo, Diana Angélica Varela-Martínez, Bárbara Socas-Rodríguez, Javier Hernández-Borges
Nanomaterials have found an important place in Analytical Chemistry and, in particular, in Separation Science. Among them, metal-organic frameworks, magnetic and non-magnetic nanoparticles, carbon nanotubes and graphene, as well as their combinations, are the most important nanomaterials that have been used up to now. Concerning capillary electromigration techniques, these nanomaterials have also been used as both pseudostationary phases in electrokinetic chromatography (EKC) and as stationary phases in microchip capillary electrophoresis (CE) and capillary electrochromatography (CEC), as a result of their interesting and particular properties...
July 6, 2017: Electrophoresis
Alexander Vladimirovich Ivanov, Polina Olegovna Bulgakova, Edward Danielevich Virus, Maria Petrovna Kruglova, Valery Vasil'evich Alexandrin, Viktoriya Aleksandrovna Gadieva, Boris Petrovich Luzyanin, Nikolai Evgen'evich Kushlinskii, Anatolij Nikolaevich Fedoseev, Aslan Amirkhanovich Kubatiev
A rapid and selective method has been developed for highly sensitive determination of total cysteine and homocysteine levels in human blood plasma and urine by capillary electrophoresis (CE) coupled with liquid-liquid extraction. Analytes were first derivatized with 1,1'-thiocarbonyldiimidazole and then samples were purified by chloroform-ACN extraction. Electrophoretic separation was performed using 0.1 M phosphate with 30 mM triethanolamine, pH 2, containing 25 μM CTAB, 2.5 μM SDS, and 2.5% polyethylene glycol 600...
July 6, 2017: Electrophoresis
Yong Deng, Shing-Chung Lam, Jing Zhao, Shao-Ping Li
Capillary zone electrophoresis was developed for the simultaneous determination of five flavonoids and one phenolic acid, including taxifolin-7-O-glucoside, flavanomarein, quercetagetin-7-O-glucoside, okanin 4'-O-glucoside, okanin and chlorogenic acid, in different parts and origins of Coreopsis tinctoria and its related species. Effects of acidity, running-buffer concentration, and modifier concentration were investigated to determine the optimum conditions for analyte determination. Analysis was performed within 18 min by using 50 mM borax buffer containing 15% acetonitrile as a modifier (pH 9...
July 6, 2017: Electrophoresis
Jun-Hui Zhang, Rui-Yu Nong, Bang-Jin Wang, Ping Ai, Sheng-Ming Xie, Li-Ming Yuan
Natural amino acids are well known to form coordination polymers with transition metal ions. In this study, six homochiral metal-organic frameworks constructed from Zn(2+) or Co(2+) ions and various enantiopure amino acid (L-tyrosine, L-histidine, L-tryptophan and L-glutamic acid), namely [Zn(L-tyr)]n (L-tyrZn), [Zn4 (btc)2 (Hbtc)(L-His)2 (H2 O)4 ]·1.5H2 O, {[Zn2 (L-trp)2 (bpe)2 (H2 O)2 ]·2H2 O·2NO3 }n , [Co2 (L-Trp)(INT)2 (H2 O)2 (ClO4 )], [Co2 (sdba)((L-Trp)2 ] and [Co(L-Glu)(H2 O)·H2 O]∞ , were synthesized according to the methods previously reported in the literature...
July 5, 2017: Electrophoresis
Sylvie Michelland, Sandrine Bourgoin-Voillard, Valérie Cunin, Axel Tollance, Pascal Bertolino, Karel Slais, Michel Seve
High-throughput mass spectrometry-based proteomic analysis requires peptide fractionation to simplify complex biological samples and increase proteome coverage. OFFGEL fractionation technology became a common method to separate peptides or proteins using isoelectric focusing in an immobilized pH gradient. However, the OFFGEL focusing process may be further optimized and controlled in terms of separation time and pI resolution. Here, we evaluated OFFGEL technology to separate peptides from different samples in the presence of low-molecular-weight (LMW) color pI markers to visualize the focusing process...
July 3, 2017: Electrophoresis
Vojtech Adam, Marketa Vaculovicova
Nanomaterials are in analytical science used for a broad range of purposes, covering the area of sample pretreatment as well as separation, detection and identification of target molecules. This part of the review covers capillary electrophoresis (CE) of nanomaterials and focuses on the application of CE as a method for characterization used during nanomaterial synthesis and modification as well as the monitoring of properties and interactions with other molecules. The heterogeneity of the nanomaterial family is extremely large...
June 30, 2017: Electrophoresis
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