• BMB Reports
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• v.39 no.2
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• pp.216-222
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• 2006

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT(dithiothreitol) and 0.2% carrier ampholytes; (b) 5M urea, 2M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N, N-dimethyl-3-ammonio-1-propanesulfonate), 40mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7M urea, 2M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.

• Applied Biological Chemistry
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• v.11
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• pp.105-108
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• 1969

본(本) 실험(實驗)에 사용(使用)된 Gel로는 Sephadex G-200, G-100과 G-75로써 fractionation은 대체(大體)로 호조건에서 이루어 졌다. G-200과 G-100의 경우에 있어서는 갈색색소의 분자크기가 서로 다른 2개(個)의 Peak를 얻을 수 있었고 Colume에 투입된 시료(試料)는 완전(完全)히 회수(回收)되었다. G-75의 경우에는 갈색색소와 분자 크기가 작은 다른 물질(物質)과를 분리(分離)할 수 있었으나 약간의 시료(試料)는 Gel에 흡착되어 완전(完全)히 회수(回收)할 수 없었다. 발효실험의 결과를 종합해 보면 우선 갈색색소를 함유하늘 모든 fraction은 발효초기에 활성(活性)을 보이며 분자크기가 작은 물질의 구획(區劃)에서는 강하고도 점증되는 활성(活性)을 관찰할 수 있었다. Sephadex Gel에 의(依)한 fractionation은 첫째로 작업(作業)도중에 시료(試料)의 변질현상(變質現象)이 없다는 것과 둘째로 갈색색소를 그들의 분자크기에 따라 분획(分劃)할 수 있다는 이점(利點)들이 있다.

• Journal of KIISE:Computing Practices and Letters
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• v.16 no.5
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• pp.551-555
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• 2010

In protein 2-DE image analysis, the accuracy of spot-matching operation which identifies the spot of the same protein in each 2-DE gel image is intensively influenced by the errors caused by the various experimental conditions. This paper proposes an efficient method to find more accurate spot-matching patterns based on multiple reference gel images in spot-matching pattern analysis in protein 2-DE image analysis. Additionally, in order to improve the reduce the execution time which is increased exponentially along with the increasing number of gel images, a "partition then extension" framework is used to find spot-matching pattern of long length and of higher accuracy. In the experiments on real 2-DE images of human liver tissue are used to confirm the accuracy and the efficiency of the proposed algorithm.

• Journal of Microbiology and Biotechnology
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• v.22 no.5
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• pp.654-658
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• 2012

Osteoarthritis (OA) is the most common rheumatic pathology. One of the major objectives of OA research is the development of early diagnostic strategies such as those using proteomic technology. Synovial fluid (SF) in OA patients is a potential source of biomarkers for OA. The efficient and reliable preparation of SF proteomes is a critical step towards biomarker discovery. In this study, we have optimized a pretreatment method for twodimensional gel electrophoresis (2DE) separation of the SF proteome, by enriching low-abundance proteins and simultaneously removing hyaluronic acid, albumin, and IgG. SF samples pretreated using this optimized method were then evaluated by 1DE and 2DE separation followed by immunodetection of cartilage oligomeric matrix protein (COMP), a known OA biomarker, and by the identification of 3 proteins (apolipoprotein, haptoglobin precursor, and fibrinogen D fragment) that are related to joint diseases.

• Journal of Microbiology and Biotechnology
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• v.16 no.1
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• pp.25-31
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• 2006

Changes of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in the glyphosate-tolerant Roundup Ready soybean were examined using purified CP4 EPSPS produced in cloned Escherichia coli as a control. CP4 EPSPS in genetically modified soybean was detected by twodimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) with databases. CP4 EPSPS in soybean products was resolved on 2-DE by first isoelectric focusing (IEF) based on its characteristic pI of 5.1, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) based on its molecular mass of 47.5 kDa. We quantified various percentages of soybean CP4 EPSPS. The quantitative analysis was performed using a 2D software program on artificial gels with spots varying in Gaussian volumes. These results suggested that 2-DE image analysis could be used for quantitative detection of GM soybean, unlike Western blotting.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.60 no.1
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• pp.29-32
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• 2015

Two-dimensional electrophoresis (2-DE) was executed to separate the seed storage proteins from the buckwheat. The proteins extracted from the whole seed proteins were better separated and observed in the use of lysis buffer. Using this method, the highly reproducible isoelectric focusing (IEF) can be obtained from polyacrylamide gels, and IEF from the polyacrylamide gel at all the possible pH range (5.0-8.0) was more easily separated than IPG (immobilized pH gradient) gels. The polyacrylamide gels in the first dimension in 2-DE was used to separate and identify a number of whole seed proteins in the proteome analysis. In this new apparatus using 2-DE, 27cm in length of plate coated with polyacrylamide gel was used and the experiment was further investigated under the various conditions.

• The korean journal of orthodontics
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• v.52 no.1
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• pp.75-79
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• 2022

Objective: To investigate remineralizing effect of three fluoride regimens on artificially demineralized enamel around orthodontic bracket by analyzing mineral density (MD) acquired from micro-computed tomography (micro-CT). Methods: Forty-eight bracket bonded bovine incisors were prepared to create demineralized enamel (DE) surface. The samples were divided into four groups according to the fluoride regimen: 1) no fluoridation, 2) 1.23% acidulated phosphate fluoride (APF) gel, 3) fluoridated toothpaste, and 4) 0.05% sodium fluoride mouthwash. Micro-CT was scanned after demineralization (T0), and 2 weeks (T1) and 4 weeks (T2) of fluoridation. Results: APF gel showed highest remineralization of DE during T1-T0 interval among the groups (p < 0.05); followed by toothpaste, mouthwash and no fluoridation. APF gel and toothpaste demonstrated significant increase in MD after 4 weeks of application (p < 0.05). Conclusions: Remineralization effects of three fluoride regimens were depicted through micro-CT analysis, of which APF gel was most effective.

• Journal of Magnetics
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• v.16 no.3
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• pp.216-219
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• 2011

Mn doped ZnO films were prepared on Si (100) substrates using sol-gel method. The prepared films were annealed at $550^{\circ}C$ for decomposition and oxidation of the precursors. XRD analysis revealed the presence of ZnMnO hexagonal wurtzite phase along with the presence of small quantity of $ZnMn_2O_3$ secondary phase and poor crystalline nature. The 2D, 3D views of magnetic domains and domain profiles were obtained using magnetic force microscopy at room temperature. Rectangular shaped domains with an average size of 4.16 nm were observed. Magnetic moment measurement as a function of magnetic field was measured using superconducting quantum interference device (SQUID) magnetometry at room temperature. The result showed the ferromagnetic hysteresis loop with a curie temperature higher than 300 K.

• Preventive Nutrition and Food Science
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• v.13 no.4
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• pp.366-369
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• 2008

Proteins secreted from bifidobacteria are believed to play important roles in human intestines via interacting with different host cells. In this respect, proteins secreted from Bifidobacterium pseudocatanulatum BP1, which has been rarely studied, were analyzed by two-dimensional gel electrophoresis (2DE). Using this approach, approx-imately 21 protein spots on a 2DE gel were detected and 10 of these spots were identified by mass spectrometry. Five spots were identified as hypothetical proteins and the remaining 5 spots were identified as a putative iron-side-rophore binding lipoprotein, a short-chain dehydrogenase/reductase SDR, an exonuclease, cytochrome P450 hydroxylase, and a putative dehydrogenase. The identification of secreted putative iron-siderophore binding lipoprotein was highly interesting since it is an important protein that is involved in ferric iron uptake in pathogenic bacteria. This finding could accelerate studies on the probiotic effect of Bifidobacterium by explaining the competition between bifidobacteria and intestinal pathogens for ferric iron.

• Proceedings of the Korean Society of Life Science Conference
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• 2002.12a
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• pp.20-47
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• 2002

Two-dimensional gel electrophoresis (2-DE) maps for human stomach tissue proteins have been prepared by displaying the protein components of the tissue by 2-DE and identifying them using mass spectrometry. This will enable us to present an overview of the proteins expressed In human stomach tissues and lays the basis for subsequent comparative proteome analysis studies with gastric diseases such as gastric cancer. In this study, 2-DE maps of soluble fraction proteins were prepared on two gel images with partially overlapping pH ranges of 4-7 and 6-9. On the gels covering pH 4-7 and pH 6-9, about 900 and 600 protein spots were detected on silver staining, respectively. For protein identification, proteins spots on micropreparative gels stained by colloidal Coomassie Brilliant Blue G-250 were excised, digested in-gel with trypsln, and analyzed by peptide mass fingerprinting with delayed extraction-matrix assisted laser dosorption/ionization-mass spectrometry (DE-MALDI-MS). In all, 243 protein spots (168 spots in acidic map and 75 spots in basic map) corresponding to 136 different proteins were identified. Besides these principal maps, maps of lower resolution, i.e. overview maps (displayed on pH 3-10 gels) for total homogenate and soluble fraction, are also presented with some identifications mapped on them. Based on the 2-DE maps presented in this study, a 2-DE database for human stomach tissue proteome has been constructed and available at http://proteome.gsnu.ac.kr/DB/2DPAGE/Stomach/. The 2-DE maps and the database resulting from this study will serve important resources for subsequent proteomic studies for analyzing the normal protein variability in healthy tissues and specific protein variations in diseased tissues.