• Title/Summary/Keyword: Gel electrophoresis

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The enhancement of protein separation by duplex SDS-PAGE (Duplex SDS-PAGE를 이용한 단백질 분리향상)

  • Pyo, Jae Sung;Roh, Si Hun;Song, Jin-Su;Lee, Kyung Hyeon;Kim, Hie-Joon;Park, Jeong Hill;Kwon, Sung Won
    • Analytical Science and Technology
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    • v.19 no.6
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    • pp.529-534
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    • 2006
  • The protein separation with molecular weight using SDS-PAGE(sodium dodecyl sulfate-polyacrylamide gel electrophoresis) is the one of the most conventional and simple techniques. In, this study, two dimensional SDS-PAGE using same separation principle consecutively was investigated and compared with one dimensional SDS-PAGE. The enhanced separation from duplex SDS-PAGE was observed and separated proteins in the gel were identified by MALDI TOF MS. Identified proteins from different gel spots were found to have different gi numbers. Therefore, duplex SDS-PAGE separation method will be used for economic separation method in the future because only tiny amount of inexpensive reagents are used to perform duplex SDS-PAGE.

The Study on the Thyroglobulin in Thyroid Diseases (갑상선질환(甲狀腺疾患)의 Thyroglobulin에 관(關)한 연구(硏究)(예보(豫報)))

  • Kim, Suk-Keun;Lim, Soo-Duk;Lee, Mun-Ho
    • The Korean Journal of Nuclear Medicine
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    • v.4 no.1
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    • pp.43-49
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    • 1970
  • In recent years with development of immuno-electrophoresis, more acurate analysis of the serum protein became possible. However, there is few reports in the literature which investigated the changes of the immunoglobulin compared with electrophoretically fractioned serum thyroglobulin in the patients with various thyroid diseases. The purpose of this report is to investigate the changes of thyroglobulin in various thyroid diseases by the method of immuno-electrophoresis and to compare the results with.serum protein fractionated by the method of agar-gel micro-electrophoresis. Materials and Methods: Sera from 9 patients with diffuse toxic goiter, 2 nodular nontoxic goiter, 2 thyroiditis, 3 hypothy, roidism, 1 thyroid cancer, 7 cystic degeneration of the thyroid gland, and 10 normal subject were taken. All cases were confirmed by various laboratory thyroid function tests and thyroid needle biopsy. Immuno-electrophoretic analysis of the serum were performed by Scheidegger's modified micro-immuno-electrophoretic method. The antiserum was obtained from the Travenol Laboratories International, Hyland Products Division and was rabbit anti-human thyroglobulin. Microscope slide agar-gel electrophoresis for serum protein fractionation was performed at $4^{\circ}C$ using veronal buffer, pH 8.6 and ionic strength 0.05, with 54 volts and 2.8 mA for 60 minutes. The fractionated slide was stained with 0.1% thiazine red. The results were as follows: 1) Increase of immune-globulin macroglobulin (IgM), alphaglobulin, and immune-globulin A (IgA) by 95.8%, 100%, 29.2% respectively was found in the serum from various thyroid diseases. 2) Thyroglobulin fraction was found to be increased in 50%, no change in 41.7%, and no line in 8.3% with all of the various goiter patients. On the other hand, 10 normal control group showed only 2 cases of increase, 5 cases of no change and 3 cases of no line.

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Purification and Characterization of Recombinant Monellin Produced from Yeast Culture Medium (효모 배양액으로부터 재조합 모넬린의 정제와 특성 연구)

  • 김인호
    • KSBB Journal
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    • v.13 no.5
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    • pp.535-539
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    • 1998
  • The monellin, a sweet-taste protein, was expressed and secreted in Saccharomyces cerevisiae. The secreted menellin was concentrated using an ultrafiltration membrane with a nominal molecular weight cut off of 3,000 or by ammonium sulfate precipitation. The monellin was purified by G-25 gel filtration chromatography, followed by CM-Sepharose ion exchange chromatography. The purified monellin was characterized by SDS-PAGE (SDS-Polyacrylamide Gel Electrophoresis) and PHLC. The molecular weight of monellin was found to be 10,700 dalton, and its purity was over 95%.

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Characterization of and Polyamine Effect on Endonuclease from Zygotes of Chlamydomonas reinhardtii (Chlamydomonas reinhardtii에서 순화한 Endonuclease의 특징과 Polyamine의 영향)

  • 김재윤
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.293-301
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    • 1990
  • We have purified and characterized a deoxyribonuclease from zygotes of the eukaryotic green alga, Chlamydomonas reinhardtii and investigated effects of the polyamines, putrescine, spermidine and spermine on the purifed endonuclease-catalyzed cleavage of plasmid DNA. The enzyme has a molecular weight of about 37 kDa as measured by gel filtration and SDS-polyacrylamide gel electrophoresis. There is no requirement for a divalent cation. The activity is sensitive to ionic strength, as NaCl and KCl result in inhibition. The cleavage of plasmid DNA by the purified endonuclease was effectively inhibited by polyamines. The enzyme activity was inhibited more effectively by spermine than by spermidine. The inhibition by putrescine was lower than the other two polyamines.

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Methodology of Genetic Engineering (遺傳工學의 方法論)

  • 이평우;윤경하;이세영
    • Korean Journal of Microbiology
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    • v.13 no.2
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    • pp.71-80
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    • 1975
  • The neurotoxin of Clostridium botulinum type B was purified from a liquid culture. The purification steps consist of ammonium sulfate precipitation of whole culture, treatment of Polymin P(0.15%, v/v), gel filtration on Sephadex G-100 at pH5.6 and DEAE-Sephadex charomatography at pH8.0. The procedure recovered 17% of the toxin assayed in the starting culture. The toxin was homogeneous by sodium dodecyl sulfate(SDS)-polyacrylamide gel electrophoresis and had a molecular weight of 163,000. Subunits of 106,000 and 56,000 molecular weight were found when purified toxin was treated with a disulfide-reducing agent and electro phoresed on SDS-polyacrylamide gels.

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Simultaneous Purification of Enterotoxin A and C by Fast Protein Liquid Chromatography (FPLC에 의한 Staphylococcal Enterotoxin A와 C의 동시분리)

  • Lee, Jung-Hee;Kim, Jong-Bae;Shin, Heuyn-Kil
    • Korean Journal of Food Science and Technology
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    • v.20 no.6
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    • pp.856-861
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    • 1988
  • A new method developed for simultaneous purification of enterotoxin A and C from Staphylococcus aureus strain L 350/1 consisted of chromatography on carboxymethyl (CM)-cellulose using a buffer of variable pH, gel filtration on Ultro gel, and fast protein liquid chromatography(FPLC) using a buffer of variable pH. The enterotoxin A and C were purified by three steps: batchwise adsorption from culture supernatant on Amberlite CG-50; chromatography on CM-cellulose using a buffer of constant pH and molarity; and gel filtration on Sephadex G-75. The purified enterotoxin appeared homogeneous by gel diffusion and polyacrylamide gel electrophoresis. Upon treatment with CM-cellulose using a elution of variable pH, enterotoxin A and C were so close that they were not separated completely. After elution from gels, the enterotoxins appeared as a single peak at the same position. Gel filtration gave a reaction of complete identity to enterotoxin A and C in Ouchterlony immunodiffusion. In FPLC using a CM-cellulose, enterotoxin A and C were simultaneously separated at pH 8.6 and 6.8. When each fraction was performed to gel immunodiffusion, at peak of enterotoxin A and C were not detected each other. In a method of elution by pH-gradient was to be more efficient as a simultaneous separation method in terms of speed, yields and simplicity. The purified toxin A and C were identical to type A and C reference enterotoxin on both disc electrophoresis and Ouchterlony gel diffusion.

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High $T_c$, Superconductors for Applications

  • Soh, Deawha;Fan, Zhanguo
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2003.05a
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    • pp.366-369
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    • 2003
  • High T$_{c}$ superconducting lines will be applied as key materials in the areas of power transmission line; magnetic levitation of vehicle; magnetic separation; magnetic energy storage and marine propulsion. A combination method of electrophoresis deposition and zone-melting for preparation of YBaCuO tape is proposed. The submicron particle powder of YBaCuO made by sol-gel method is used in the electrophoresis process. A 40~50${\mu}{\textrm}{m}$ thickness of YBaCuO film on Ag plate could be deposited in about three minutes. After deposition the film is rolled and heat treated in order to increase the density and the adhesion of the film to the Ag plate. Silver(Ag) and lead oxide(PbO) were added in the YBaCuO powder in order to reduce its melting point. The YBaCuO coating with controlled Ag and PbO contents was preliminarily zone-melted at about 945$^{\circ}C$.>.

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Analysis of Double-Stranded DNA Fragments by Capillary Electrophoresis Using Entangle Polymer Solutions in Uncoated Fused Silica Capillary Columns

  • Lee, Jong-Jin;Lee, Kong-Joo
    • BMB Reports
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    • v.31 no.4
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    • pp.384-390
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    • 1998
  • DNA fragments (51-587 bp) were separated by capillary electrophoresis using entangled polymer, hydroxyethylcellulose, in uncoated fused silica capillary columns. The factors affecting the separation of DNA fragments with hydroxyethylcellulose media were evaluated, i.e., the concentration of buffer and entangled polymer, effects of additives (methanol, ethidium bromide, EDTA), temperature, and injection methods. Maximum performance was obtained by adding 5% methanol in 0.5% hydroxyethylcellulose solution at $30^{\circ}C$. Addition of methanol in polymer media increased the resolution of small size DNA fragments (< 100 bp). On the other hand, addition of ethidium bromide and EDTA, which are commonly used in conventional DNA separation, reduced the resolution of DNA fragments in the polymer solution. It turns out that the separation behavior of DNA in entangled polymer is more sensitive to the running condition compared to that in polyacrylamide gel-filled capillary, but the reproducibility of DNA separation in entangled polymer is reliable.

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