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Protein electrophoresis

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https://www.readbyqxmd.com/read/28812992/endolysosomal-degradation-of-allergenic-ole-e-1-like-proteins-analysis-of-proteolytic-cleavage-sites-revealing-t-cell-epitope-containing-peptides
#1
Sabrina Wildner, Brigitta Elsässer, Teresa Stemeseder, Peter Briza, Wai Tuck Soh, Mayte Villalba, Jonas Lidholm, Hans Brandstetter, Gabriele Gadermaier
Knowledge of the susceptibility of proteins to endolysosomal proteases provides valuable information on immunogenicity. Though Ole e 1-like proteins are considered relevant allergens, little is known about their immunogenic properties and T cell epitopes. Thus, six representative molecules, i.e., Ole e 1, Fra e 1, Sal k 5, Che a 1, Phl p 11 and Pla l 1, were investigated. Endolysosomal degradation and peptide generation were simulated using microsomal fractions of JAWS II dendritic cells. Kinetics and peptide patterns were evaluated by gel electrophoresis and mass spectrometry...
August 16, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28812359/continuous-flow-electrophoresis-of-dna-and-proteins-in-a-two-dimensional-capillary-well-sieve
#2
Lian Duan, Zhen Cao, Levent Yobas
Continuous-flow electrophoresis of macromolecules is demonstrated using an integrated capillary-well sieve arranged into a two-dimensional anisotropic array on silicon. The periodic array features thousands of entropic barriers, each resulting from an abrupt interface between a 2-μm-deep well (channel) and a 70-nm capillary. These entropic barriers owing to two-dimensional confinement within the capillaries are vastly steep in relation to those arising from slits featuring one-dimensional confinement. Thus, the sieving mechanisms can sustain relatively large electric field strengths over a relatively small array area...
August 16, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28811086/proteomic-profiling-of-the-antifungal-drug-response-of-aspergillus-fumigatus-to-voriconazole
#3
Nansalmaa Amarsaikhan, Daniela Albrecht-Eckardt, Christoph Sasse, Gerhard H Braus, Zumrut B Ogel, Olaf Kniemeyer
Antifungal resistance is an emerging problem and one of the reasons for treatment failure of invasive aspergillosis (IA). Voriconazole has become a standard therapeutic for the treatment of this often fatal infection. We studied the differentially expressed proteins as a response of Aspergillus fumigatus to voriconazole by employing the two-dimensional difference gel electrophoresis (DIGE) technique. Due to addition of drug, a total of 135 differentially synthesized proteins were identified by MALDI-TOF/TOF-mass spectrometry...
August 5, 2017: International Journal of Medical Microbiology: IJMM
https://www.readbyqxmd.com/read/28809090/the-association-between-p-selectin-glycoprotein-ligand-1-gene-variable-number-of-tandem-repeats-polymorphism-and-risk-of-thrombosis-in-beh%C3%A3-et-s-disease
#4
Fulya Cosan, Basar Oku, Ozgun M Gedar Totuk, Neslihan Abaci, Duran Ustek, Reyhan Diz Kucukkaya, Ahmet Gul
OBJECTIVES: Behçet's disease (BD) has been recognized as an unclassified type of vasculitis with an accompanying tendency to thrombosis. No disease-specific pathology has been demonstrated so far to explain the prothrombotic state, and this predisposition is considered to be associated with endothelial activation/dysfunction. P-selectin glycoprotein ligand-1 (PSGL-1) variable number of tandem repeat (VNTR) polymorphism has an impact on the protein length, and heterozygosity affect of the PSGL-1 to P-selectin interaction, which has been found to be associated with an increased risk of thrombosis in patients with antiphospholipid syndrome...
August 14, 2017: International Journal of Rheumatic Diseases
https://www.readbyqxmd.com/read/28808999/studying-protein-protein-interactions-by-biotin-ap-tagged-pulldown-and-ltq-orbitrap-mass-spectrometry
#5
Zhongqiu Xie, Yuemeng Jia, Hui Li
The study of protein-protein interactions represents a key aspect of biological research. Identifying unknown protein binding partners using mass spectrometry (MS)-based proteomics has evolved into an indispensable strategy in drug discovery. The classic approach of immunoprecipitation with specific antibodies against the proteins of interest has limitations, such as the need for immunoprecipitation-qualified antibody. The biotin AP-tag pull-down system has the advantage of high specificity, ease of use, and no requirement for antibody...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28808482/the-chinese-herb-prunella-vulgaris-promotes-apoptosis-in-human-well-differentiated-thyroid-carcinoma-cells-via-the-b-cell-lymphoma-2-bcl-2-associated-x-protein-caspase-3-signaling-pathway
#6
De-Tao Yin, Mengyuan Lei, Jianhui Xu, Hongqiang Li, Yongfei Wang, Zhen Liu, Runsheng Ma, Kun Yu, Xianghua Li
Prunella vulgaris (PV), a traditional Chinese herb, has been shown to be rich in bioactive chemicals and possess anti-proliferative and pro-apoptotic effects on tumor cells. The effect of PV on human well-differentiated thyroid carcinoma (WDTC), which accounts for the majority of common endocrine malignancies, remains to be elucidated. The present study aimed to investigate the function of PV on WDTC cell lines and apoptosis-associated signaling pathway activity. Additional studies demonstrated that PV may induce apoptosis in WDTC TPC-1 and FTC-133 cell lines, using the Cell Counting Kit-8 assay...
August 2017: Oncology Letters
https://www.readbyqxmd.com/read/28806997/stathmin-is-overexpressed-and-regulated-by-mutant-p53-in-oral-squamous-cell-carcinoma
#7
Hai-Long Ma, Shu-Fang Jin, Wu-Tong Ju, Yong Fu, Yao-Yao Tu, Li-Zhen Wang, Jiang-Li, Zhi-Yuan Zhang, Lai-Ping Zhong
BACKGROUND: The aim of this study was to investigate the oncogenic function and regulatory mechanism of stathmin in oral squamous cell carcinoma (OSCC). METHODS: Two-dimensional electrophoresis and liquid chromatography-tandem mass chromatography were applied to screen differentiated proteins during carcinogenesis in OSCC. Cell Counting Kit-8 (CCK-8) assays, colony formation, migration, flow cytometry, immunofluorescence and a xenograft model were used to detect the function of stathmin...
August 14, 2017: Journal of Experimental & Clinical Cancer Research: CR
https://www.readbyqxmd.com/read/28805727/leaf-physiological-and-proteomic-analysis-to-elucidate-silicon-induced-adaptive-response-under-salt-stress-in-rosa-hybrida-rock-fire
#8
Prabhakaran Soundararajan, Abinaya Manivannan, Chung Ho Ko, Sowbiya Muneer, Byoung Ryong Jeong
Beneficial effects of silicon (Si) on growth and development have been witnessed in several plants. Nevertheless, studies on roses are merely reported. Therefore, the present investigation was carried out to illustrate the impact of Si on photosynthesis, antioxidant defense and leaf proteome of rose under salinity stress. In vitro-grown, acclimatized Rosa hybrida 'Rock Fire' were hydroponically treated with four treatments, such as control, Si (1.8 mM), NaCl (50 mM), and Si+NaCl. After 15 days, the consequences of salinity stress and the response of Si addition were analyzed...
August 14, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28804967/comparative-proteomic-analysis-of-different-developmental-stages-of-swamp-buffalo-testicular-seminiferous-tubules
#9
P-F Zhang, Y-L Huang, Q Fu, F-M Chen, Y-Q Lu, X-W Liang, M Zhang
With ageing, many protein components change markedly during mammalian spermatogenesis. Most of these proteins have yet to be characterized and verified. Here, we have employed two-dimensional electrophoresis coupled to tandem mass spectrometry to explore the different proteins from pre-pubertal, pubertal and post-pubertal swamp buffalo testicular seminiferous tubules. The results showed that 25 protein spots were differentially expressed among developmental stages, and 13 of them were successfully identified by mass spectrometry...
August 13, 2017: Reproduction in Domestic Animals, Zuchthygiene
https://www.readbyqxmd.com/read/28804850/laboratory-testing-for-von-willebrand-factor-multimers
#10
Susan Oliver, Kun Kan Edwin Lau, Kent Chapman, Emmanuel J Favaloro
Von Willebrand disease (VWD) is reportedly the most common inherited bleeding disorder and can also arise as an acquired syndrome (AVWS). These disorders develop due to defects and/or deficiency of the plasma protein von Willebrand factor (VWF). Laboratory testing for the VWF-related disorders requires assessment of both VWF level and VWF activity, the latter requiring multiple assays because of the many functions carried out by VWF to help prevent bleeding. As an additional step, an evaluation of VWF structural features by multimer analysis is useful in selective investigations...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28803590/differential-protein-analysis-of-serum-exosomes-post-intravenous-immunoglobulin-therapy-in-patients-with-kawasaki-disease
#11
Li Zhang, Qi-Fang Song, Jing-Jie Jin, Ping Huang, Zhou-Ping Wang, Xiao-Fei Xie, Xiao-Qiong Gu, Xue-Juan Gao, Hong-Ling Jia
BACKGROUND: Kawasaki disease, which is characterised by systemic vasculitides accompanied by acute fever, is regularly treated by intravenous immunoglobulin to avoid lesion formation in the coronary artery; however, the mechanism of intravenous immunoglobulin therapy is unclear. Hence, we aimed to analyse the global expression profile of serum exosomal proteins before and after administering intravenous immunoglobulin. METHODS: Two-dimensional electrophoresis coupled with mass spectrometry analysis was used to identify the differentially expressed proteome of serum exosomes in patients with Kawasaki disease before and after intravenous immunoglobulin therapy...
August 14, 2017: Cardiology in the Young
https://www.readbyqxmd.com/read/28803541/ion-exchange-chromatography-purification-of-extracellular-vesicles
#12
Maja Kosanović, Bojana Milutinović, Sanja Goč, Ninoslav Mitić, Miroslava Janković
Despite numerous studies, isolating pure preparations of extracellular vesicles (EVs) has proven challenging. Here, we compared ion-exchange chromatography (IEC) to the widely used sucrose density gradient (SDG) centrifugation method for the purification of EVs. EVs in bulk were isolated from pooled normal human amniotic fluid (AF) by differential centrifugation followed by IEC or sucrose density gradient separation. The purity of the isolated EVs was evaluated by electrophoresis and lectin blotting/immuno blotting to monitor the distribution of total proteins, different EVs markers, and selected N-glycans...
August 1, 2017: BioTechniques
https://www.readbyqxmd.com/read/28803268/proteomic-profiling-of-the-dystrophin-complex-and-membrane-fraction-from-dystrophic-mdx-muscle-reveals-decreases-in-the-cytolinker-desmoglein-and-increases-in-the-extracellular-matrix-stabilizers-biglycan-and-fibronectin
#13
Sandra Murphy, Heinrich Brinkmeier, Mirjam Krautwald, Michael Henry, Paula Meleady, Kay Ohlendieck
The almost complete loss of the membrane cytoskeletal protein dystrophin and concomitant drastic reduction in dystrophin-associated glycoproteins are the underlying mechanisms of the highly progressive neuromuscular disorder Duchenne muscular dystrophy. In order to identify new potential binding partners of dystrophin or proteins in close proximity to the sarcolemmal dystrophin complex, proteomic profiling of the isolated dystrophin-glycoprotein complex was carried out. Subcellular membrane fractionation and detergent solubilisation, in combination with ion exchange, lectin chromatography and density gradient ultracentrifugation, was performed to isolate a dystrophin complex-enriched fraction...
August 12, 2017: Journal of Muscle Research and Cell Motility
https://www.readbyqxmd.com/read/28802807/acute-phase-response-in-babesia-canis-and-dirofilaria-immitis-co-infections-in-dogs
#14
Zorana Milanović, Anja Ilić, Jelena Francuski Andrić, Vladimir Radonjić, Anđelo Beletić, Milica Kovačević Filipović
Babesia canis and Dirofilaria immitis are emerging and geographically overlapping vector-borne pathogens in dogs. Infection with B. canis leads to acute-phase response (APR) that can be mild to severe and results in either non-complicated or complicated forms of the disease. The aim of this study was to determine whether acute B. canis infection is more severe in dogs with underlying asymptomatic D. immitis infection. Dogs of both sexes, different ages and breeds, with naturally occurring mono-infections with B...
August 2, 2017: Ticks and Tick-borne Diseases
https://www.readbyqxmd.com/read/28802310/quantification-of-whey-protein-content-in-infant-formulas-by-sodium-dodecyl-sulfate-capillary-gel-electrophoresis-sds-cge-single-laboratory-validation-first-action-2016-15
#15
Ping Feng, Christophe Fuerer, Adrienne McMahon, Darryl Sullivan, John Austad, Sneh Bhandari, Esther Campos-Gimenez, Scott Christiansen, Hans Cruijsen, Jon DeVries, Brendon Gill, Don Gilliland, Min Huang, Estela Kneeteman, Adrienne McMahon, Bill Mindak, Maria Ofitserova, Melissa Phillips, Shay Phillips, Karen Schimpf, Guenther Raffler, Kate Rimmer, Wil van Loon, Jinchuan Yang
No abstract text is available yet for this article.
July 1, 2017: Journal of AOAC International
https://www.readbyqxmd.com/read/28801044/cationic-ionic-liquid-surfactant-polyacrylamide-gel-electrophoresis-for-enhanced-separation-of-acidic-and-basic-proteins-with-single-step-ribonuclease-b-glycoforms-separation
#16
Punprabhashi Vidanapathirana, Farhana Hasan, Kaitlyn Mussio, Anuja Pande, Michael Brands, Noureen Siraj, Anne Grove, Isiah M Warner
Cationic ionic liquids-based surfactants (ILS), such as 4-methyl pyridinium bromide (CnPBr, where n=4,6,8), were used in preparation of polyacrylamide gels, sample buffer, and running buffer for cationic ILS polyacrylamide gel electrophoresis (ILS-PAGE). These ILS are liquids in the pure state and were selected for improved separation of ribonuclease b (Rib b) glycoforms in a single step and a protein mixture containing bovine serum albumin (BSA, pI-4.8, 66.5kDa), ovalbumin (Ova, pI-4.6, 44.3kDa), α-chymotrypsinogen (α-Chy, pI-8...
July 29, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28800846/saprotrophic-proteomes-of-biotypes-of-the-witches-broom-pathogen-moniliophthora-perniciosa
#17
Sandra Pierre, Gareth W Griffith, Russell M Morphew, Luis A J Mur, Ian M Scott
Nine geographically diverse Moniliophthora perniciosa (witches' broom disease pathogen) isolates were cultured in vitro. They included six C-biotypes differing in virulence on cacao (Theobroma cacao), two S-biotypes (solanaceous hosts), and an L-biotype (liana hosts). Mycelial growth rates and morphologies differed considerably, but no characters were observed to correlate with virulence or biotype. In plant inoculations using basidiospores, one C-biotype caused symptoms on tomato (an S-biotype host), adding to evidence of limited host adaptation in these biotypes...
September 2017: Fungal Biology
https://www.readbyqxmd.com/read/28800544/protease-loaded-permeation-enhancer-liposomes-for-treatment-of-skin-fibrosis-arisen-from-second-degree-burn
#18
Kamini Sahu, Monika Kaurav, Ravi Shankar Pandey
Cysteine protease (papain) is a plant derived enzyme and due to its collagenolytic activity has potential in fibrosis reduction. However, a major hurdle in its use as fibrosis reducing agent is to overcome stratum corneum skin barrier via topical application, owing to its hydrophilic and high molecular weight and protein nature which is prone to degradation. The aim of the present study was to develop a penetration enhancer incorporated drug delivery system, i.e. propylene glycol (PG) liposomes, loaded with papain for application in fibrosis therapy...
August 8, 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/28800361/identification-of-a-common-ara-h-3-epitope-recognized-by-both-the-capture-and-the-detection-monoclonal-antibodies-in-an-elisa-detection-kit
#19
Lipei Zhao, Liang Zhao, Buchang Zhang, Jason M Robotham, Kenneth H Roux, Hengli Tang
Allergy to peanuts has become a common and severe problem, especially in westernized countries. In this study, we evaluated the target and epitope specificity of the capture and detection mouse monoclonal antibodies (mAbs) used in a commercial peanut allergen detection platform. We first identified the target of these antibodies as Ara h 3 and then used an overlapping peptide array of Ara h 3 to determine the antibody-binding epitopes. Further amino acids critical for the binding via alanine substitutions at individual amino acid residues within the epitope were mapped...
2017: PloS One
https://www.readbyqxmd.com/read/28799534/cutaneous-plasmacytosis-a-rare-entity-with-unique-presentation
#20
Subhra Dhar, Lalthleng Liani, Kamlakar Patole, Sandipan Dhar
Primary cutaneous plasmacytosis is a rare cutaneous disorder with extensive cutaneous plaques/papules mainly on the trunk and face. Cases have mostly been documented from Japan. We present here a rare case of cutaneous plasmacytosis from India of Mongolian descent. This 50-year-old female from Mizoram had extensive maculo-papular violaceous plaques distributed on the face, axillae, trunk and lower extremities. Initial and repeat skin biopsy revealed dense perivascular and periadnexal mature plasma cells. She also had lymphadenopathy...
August 9, 2017: Indian Journal of Dermatology, Venereology and Leprology
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