• Title/Summary/Keyword: SDS electrophoresis analysis

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Analysis of Branched PEG-Conjugated Interferon Alpha by Capillary Electrophoresis and MALDI- TOF Mass Spectrometry

  • Na, Dong-Hee;Park, Eun-Ji;Lee, Snag-Deuk;Jo, Young-Woo;Lee, Sung-Hee;Kim, Won-Bae;Lee, Kang-Choon
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.406.3-407
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    • 2002
  • Analysis of interferon alpha (IFN) modified with high molecular weight branched PEG was performed by capillary electrophoresis (CE) and MALDI-TOF mass spectrometry (MALDI-TOF MS). IFN was modified by the reaction of amine residues with an active ester of monomethoxy polyethylene glycol at various molar ratios. As a CE method. capilary electrophoresis sodium dodecyl sulfate nongel sieving (CE-SDS NGS) was performed using an uncoated capilary filled with a hydrophilic replaceable polymer network matrix. (omitted)

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Classification of Meat-Based Listeria monocytogenes Using Whole-Cell Protein Patterns and Serotyping Analysis

  • Park, Si-Hong;Jung, Sang-Hoon;Kim, Hyun-Joong;Chung, Yun-Hee;Kim, Hae-Yeong
    • Food Science and Biotechnology
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    • v.15 no.2
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    • pp.324-327
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    • 2006
  • The food-borne pathogen Listeria monocytogenes is commonly associated with meats and unpasteurized dairy products. To identify this pathogen in meats more efficiently than has been done in the past, we purchased meats from Korean markets and performed sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and serotyping analysis on Listeria organisms isolated from meat samples. Each Listeria species showed specific protein band patterns on SDS-PAGE. Whole-cell protein SDS-PAGE profiles indicated that the organisms isolated from meats sold in local Korean markets were L. monocytogenes with the serotypes 1/2a, 1/2b, 1/2c, and 4b. We suggest that it is possible to carry out molecular subtyping of L. monocytogenes using SDS-PAGE.

Quantitation of CP4 5-Enolpyruvylshikimate-3-Phosphate Synthase in Soybean by Two-Dimensional Gel Electrophoresis

  • KIM YEON-HEE;CHOI SEUNG JUN;LEE HYUN-AH;MOON TAE WHA
    • 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.

Proteome Analysis of Responses to Ascochlorin in LPS-induced Mouse Macrophage RAW264.7 Cells by 2-D Gel Electrophoresis and MALDI-TOF MS. (LPS로 자극된 macrophage RAW264.7 세포에서 ascochlorin에 대한 단백질체 분석)

  • Chang, Young-Chae
    • Journal of Life Science
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    • v.18 no.6
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    • pp.814-825
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    • 2008
  • Ascochlorin (ASC) is prenyl-phenol compound that was isolated from the fungus Ascochyta viciae. ASC reduces serum cholesterol and triglyceride levels, and suppresses hypertension, tumor development, ameliorates type I and II diabetes. Here, to better understand the mechanisms by which ASC regulates physiological or pathological events and induces responses in the pharmacological treatment of inflammation, we performed differential analysis of the proteome of the mouse macrophage RAW264.7 cells in response to ASC. In this study, we used a proteomic analysis of LPS-induced RAW264.7 cells treated by ASC, to identify proteins potentially involved in inflammatory processes. The RAW264.7 cell proteomes with and without treatment with ASC were compared using two-dimensional electrophoresis (2-D SDS-PAGE), matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS) and bioinformatics. The largest differences in expression were observed for the calreticulin (4-fold decrease), ${\beta}-actin$ (4-fold decrease) and vimentin (1.5-fold decrease). In addition, rabaptin was increased 3-fold in RAW264.7 cells treated with ASC. The expression of some selected proteins was confirmed by RT-PCR analysis.

Presence of an Albumin-Like Protein on the Plasma Membrane of Adipocytes Isolated from Korean Cow

  • Choi, Chang-Bon;Rhee, Seung-Keun
    • BMB Reports
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    • v.31 no.6
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    • pp.542-547
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    • 1998
  • The main function of adipocytes in various species is to store nutrient energy in the form of triglycerides, and this function may he closely related with hormonal signaling through the plasma membrane of adipocytes. Using SDS-PAGE, two-dimensional gel electrophoresis, and a membrane biotinylation technique, we have identified a 55KDa protein (55K protein) from the plasma membrane fraction of adipocytes, with an isoelectric point (pI) of 8.1-8.3. However, this 55K protein was not observed with a two-dimensional gel electrophoresis carried out on plasma membrane fractions prepared from the liver, heart, and kidney tissues. An analysis of the 12 amino acids sequence at the N-terminal of the 55K protein showed that it has a similar sequence to that of bovine serum albumin.

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Effectiveness for Beauty Improvement of Geranium essential oil on Skin Troubles by Surfactant (계면활성제에 의해 손상된 피부의 Geranium essential oil 미용증진 효과)

  • Choi, Jeung-Suk
    • Journal of the Korean Society of Fashion and Beauty
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    • v.2 no.2 s.2
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    • pp.101-106
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    • 2004
  • Allergic contact dermatitis is a common skin disease resulting from specific immunologic sensitization to topically applied various allergen. The Purpose of this study was to investigate skin morphologic cahnges by light microscopic and scanning electron Microscope, changes of protein band by SDS-PAGE(SDS-polyacrylamide gel electrophoresis) in the skin effectiveness for beruty improvement of Geranium essential oil on skin troubles by surfactant. The results of the study are as follows, 1. From the observed result of FE-Scanning Electron Microscope, groups treated by Geranium essential oil in group treated Geranium essential oil during 1 week in surfactant treated group during 1 week, the group was repaired in irregularity surface of tissue by alleviate-keratinization of Geranium essential oil. 2. As a result of protein analysis the group treated on surfactant was rised protein upper range of 97,0004a11on by hyper-keratinization and group treated during 1 week by surfactant was decreased protein below range of 43,000dalton.

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Comparative antigen analysis of yrichomonus vaginulis by enzyme-linked immunoelectrotransfer blot technique (효소면역 전기영동 이적법을 이용한 질트리코모나스 항원의 비교 분석)

  • 민득영;임미혜
    • Parasites, Hosts and Diseases
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    • v.30 no.4
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    • pp.323-328
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    • 1992
  • Analysis of siR isolates of Trichemenes veginalis was carried out with sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunoelectrotransfer blot (EITB). Trichloroacetic acid-treated antigens of the 6 isolates revealed 25 protein profiles ranging 12~170 kDa of molecular weight in SDS-PAGE. In EITB, the specific immunogenic bands were visualized at 51 kDa and 96 kDa when HY-1 antigen was probed with difFerent mice sera immunized with 6 isolates of T. vaginalis. The banding patterns with different sera showed isolate-to-isolate variability. In EITB, homologous antigen (HY-1) did not show any enhanced response in reacting to homologous antiserum(HY-1) when 6 isolates of T. vaginalis were probed with a single serum (HY-1). It is assumed that the different banding patterns of six isolates show isolate-to-isolate variability and immunogenic common bands in 41, 47, 74 and 9:k kDa on EITB may connote the important significance on immune response in T. vaginalis infection.

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Soluble Proteins Analysis of Class Cephalopoda in the Yellow Sea(I) (황해산 두족류의 가용성 단백질에 대한 연구 (I))

  • 허회권
    • Journal of Aquaculture
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    • v.10 no.3
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    • pp.301-310
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    • 1997
  • To investigate a possibility of the species genetic relationship for the soluble proteins analysis on the class Cephalopoda in the Yellow Sea, the isolate eye, muscle and liver proteins from five species (Sepia esculenta, Sepiella japonica, Loligo chinensis, Loligo beka and Octopus minor) were analysed using different electrophoretic techniques (Davis-polyacrylamide gel electrophoresis, SDS-PAGE, exponential gradient SDS-PAGE, thin-layer isoelectro-focusing and two-dimensional PAGE). The average molecular weight of the soluble eye and muscle proteins was estimated at 35-50 KDa, separated b the exponential gradient SDS-PAGE. It was corresponds to that of electrophoretic patterns by t재 dimensional PAGE. By which the thin layer IEF, the target proteins showed a reasonable specificity based on their isoelectric points (pI) 7.5-8.5.

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Studies on the Transformation of Crop Plants. IV. Biochemical Characteristics of Embryogenic Callus in Rice (곡물류의 형질전환 유도에 관한 연구 (IV) 벼 배발생 세포의 생화학적 특징)

  • 정병균
    • Journal of Plant Biology
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    • v.36 no.4
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    • pp.377-382
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    • 1993
  • Rice (Oryza saliva L.) calli containing both embryogenic callus (EC) and non embryogenic callus (NEC) regions were initiated from the mature seed on MS medium supplemented with 2.0 mg/L 2,4-D, 0.5 mg/L kinetin. The calli were developed into two callus type which can be distinguished by visual examination depending on color and appearance. In order to illucidate the polypeptide composition between EC and NEC, the total protein extracted from two types of callus was analysed by electrophoresis. By one-dimesional anlaysis of SDS-PAGE and Isoelectric focusing, several protein bands showed quantitative and qualitative difference in each type of callus. The further analysis of the total protein with two-dimensional electrophoresis showed at least 20 EC specific protein and 10 NE specific protein. Also 3 specific protein spots showing micro heterogeneity of 90, 65, 50 kD were detected in EC, while a series of acidic heterologous protein spots were visualized in NEC.in NEC.

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Proteomic Assessment of Dung Beetle, Copris tripartitus Immune Response

  • Suh, Hwa-Jin;Bang, Hea-Son;Kim, Seong-Ryul;Yun, Eun-Young;Park, Kwan-Ho;Kang, Bo-Ram;Kim, Ik-Soo;Jeon, Jae-Pil;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.17 no.2
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    • pp.217-221
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    • 2008
  • Dung beetle larvae at the $3^{rd}$ instar were injected with lipopolysaccaride and inducible proteins were examined within a pI level of 3-10 and a size level by proteomics, including 1-D SDS PAGE analysis and antibacterial assay. The immune infected larvae extracts provided seven protein bands in one-dimensional electrophoresis and its antibacterial activity also checked. Hemolymph protein from immune infected larvae of the dung beetle were separated by twodimensional gel electrophoresis and compared with those from native larvae. In 2-D gel electrophoresis, we detected 63 immune infected unique and 32 up-regulated proteins, and 36 proteins that were down-regulated or not present in treated gel. Ten protein spots from unique proteins and those presented as different level of abundance in infected and native larvae were specially expressed. These differentially expressed proteins were proposed to be involved in the defense mechanism against microorganism.