• Title/Summary/Keyword: Electrophoresis

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Affinity chromatography and capillary electrophoresis for analysis of the yeast ribosomal proteins

  • Goyder, Miriam S.;Willison, Keith R.;Klug, David R.;DeMello, Andrew J.;Ces, Oscar
    • BMB Reports
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    • v.45 no.4
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    • pp.233-238
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    • 2012
  • We present a top down separation platform for yeast ribosomal proteins using affinity chromatography and capillary electrophoresis which is designed to allow deposition of proteins onto a substrate. FLAG tagged ribosomes were affinity purified, and rRNA acid precipitation was performed on the ribosomes followed by capillary electrophoresis to separate the ribosomal proteins. Over 26 peaks were detected with excellent reproducibility (<0.5% RSD migration time). This is the first reported separation of eukaryotic ribosomal proteins using capillary electrophoresis. The two stages in this workflow, affinity chromatography and capillary electrophoresis, share the advantages that they are fast, flexible and have small sample requirements in comparison to more commonly used techniques. This method is a remarkably quick route from cell to separation that has the potential to be coupled to high throughput readout platforms for studies of the ribosomal proteome.

The Enhanced Electrophoresis Method in Leachate System for Repairing of Leaks in Waste Landfill Geomembrane Liner (폐기물 매립지 차수층 누출시 전기영동 복원을 위한 침출수에서의 향상기법)

  • Kim, Jong-Yun;Han, Sang-Jae;Kim, Soo-Sam
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.30 no.1C
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    • pp.7-15
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    • 2010
  • In case that the seepage of contaminants into the subsurface has been generated from the waste impoundment by demage of geomembrane liner, it is necessary to repair the leaks of geomembrane liner for minimizing the environmental contamination by electrophoresis method. However, when electrophoresis method is applied to leachate electrolyte system, the phenomenon of clay particles flocculation would be accelerated by the interaction between clay particles and specific chemicals in leachate. In addition, the gravitational settling behaviour would be induced superior to the electrophoretic migration behaviour. Eventually, the limitations of field applicability for using the electrophoresis method are appeared. Therefore, 1-D enhanced electrophoresis method is conducted to prevent the clay flocculation and accelerate the migration of clay particles separately. After the 1-D enhanced electrophoresis experiment, we can get the results that the deflocculation effect of clay particles is increased by electrical repulsion of polymer, which adsorbed in clay particle edge, in case of using PAA dispersing agent.

Studies on the Identification of Turfgrass by Electrophoresis (SDS-PAGE, PAGIF) (전기영동법 (SDS-PAGE, PAGIF)에 의한 잔디 분류에 관한 연구)

  • 박재복;김영후;이수영
    • Asian Journal of Turfgrass Science
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    • v.5 no.1
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    • pp.11-22
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    • 1991
  • This experiment was executed to investigate the possibility of the application of taxonomic method through the isoelectric focusing with polyacrylamide gel and sodium dodecyl sulfate-polyacrylamide gel electrophoresis with seeds in the identification of turfgrasses. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to investigate the pattern of seed proteins which were extracted from 18 cultivars of cool season turfgrass and 4 cultivars of warm season turfgrass. The isoelectric focusing with polyacrylarnide gel was used to investigate the activity of the three isozymes of esterase, peroxidase and phosphoglucose isomerase which were extracted from 18 cultivars of cool season turfgrass and 4 cultivars of warm season turfgrass. The results were summarized as follows. 1. The difference of the patterns of seed proteins was observed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The identification of intra-genus was easily detected. 2. The three isozymes of esterase, peroxidase and phosphoglucose isomerase were investigated through isoelectric focusing with polyacrylamide gel. As a result, esterase was most effective among three isozymes in the identification of turfgrass cultivars 3. In the past cultivar identification was primarily based on visual morphological characters, but there was a lot of difficulty. If we should use electrophoresis, we will be able to identifvturfgrass cultivars more effectively.

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Preparation of Agarose from Gelidium amansii for Gel Electrophoresis using Various Purification Methods and Its Resolution Characteristics for DNA (다양한 정제방법에 의한 전기영동용 한천유래 아가로즈의 제조 및 DNA분리 특성)

  • Do, Jeong-Ryong;Oh, Se-Wook
    • Korean Journal of Food Science and Technology
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    • v.31 no.1
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    • pp.110-114
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    • 1999
  • The present study was conducted to investigate the preparative methods of agarose for gel electrophoresis from agar. Naturally occuring agar consists of two main polysaccharides, the neutral polysaccharide agarose and the acid sulphated polysaccharide agaropectin. The sulphate and carboxyl functions of the agar are accumulated in the agaropectin. The hydrophilic, non-ionogenic, rigid and transparent gel matrix of the agarose was found to be suitable for gel electrophoresis gel filtration and affinity chromatography. Agar was purified by chitosan treatment, cetylpyridinium chloride (CPC) treatment, and polyethylene glycol (PEG) treatment. Yields of agarose purified from agar with chitosan, CPC and PEG were 56.7%, 55.6% and 62.3%. It was proper to treat with chitosan in preparative methods of agarose for gel electrophoresis from agar.

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Purification of Peptide Components including Melittin from Bee Venom using gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis (Gel filtration chromatography와 propionic acid/urea polyacrylamide gel electrophoresis를 이용한 봉독 성분의 분리)

  • Choi, Young-Chon;Choi, Suk-Ho;Kwon, Ki-Rok
    • Journal of Pharmacopuncture
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    • v.9 no.2
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    • pp.105-111
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    • 2006
  • Objectives : This study was conducted to carry out Purification of Melittin and other peptide components from Bee Venom using gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis Methods : Melittin and other peptide components were separated from bee venom by using gel filtration chromatography on Sephadex G-50 column in 0.05M ammonium acetate buffer. Results : Melittin and other peptide components were separated from bee venom by using gel filtration chromatography on Sephadex G-50 column in 0.05M ammonium acetate buffer. The fractions obtained from gel filtration chromatography was analyzed by using SDS-PAGE and propionic acid/urea polyacrylamide gel electrophoresis. The melittin obtained from the gel filtration contained residual amount of phospholipase $A_2$ and a protein with molecular weight of 6,000. The contaminating proteins were removed by the second gel filtration chromatography. Conclusion : Gel filtration chromatography and propionic acid/urea polyacrylamide gel electrophoresis are useful to separate peptide components including melittin from bee venom.

Formation of MgO Thick Film Layer for AC-PDP via Electrophoresis Deposition of Nano-sized MgO Powders

  • Ko, Min-Soo;Kim, Yong-Seog
    • Journal of Information Display
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    • v.8 no.2
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    • pp.25-31
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    • 2007
  • MgO thick film for ac-PDPs was formed via electrophoresis deposition process and its effect on luminance and luminance efficiency were evaluated. The electrophoresis deposition process of MgO thick film was optimized through parametric study and defects levels in MgO powders was evaluated using cathodoluminescence spectra measurements. The results demonstrate a possibility of using MgO thick film as electron emission layer for ac-PDPs.

Analysis of Vaginal Lactic Acid Producing Bacteria in Healthy Women

  • Nam, Hye-Ran;Whang, Kyung-Hee;Lee, Yeon-Hee
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.515-520
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    • 2007
  • Vaginal lactic acid-producing bacteria of 80 pre-menopausal women were studied by isolation on Blood and DeMan-Rogosa-Sharpe agar, PCR with group-specific primers for Lactobacillus-denaturing gradient gel electrophoresis (DGGE), and PCR with specific primers for V3 region in 16S rRNA-temporal temperature gel electrophoresis (TTGE). Conventional isolation method on media detected only one lactobacillus (Lactobacillus brevis) while TTGE detected only Lactobacillus sp. DGGE detected seven Lactobacillus species; L. coleohominis, L. crispatus, L. iners, L. reuteri, L. rhamnosus, L. vaginalis, and Leuconostoc lactis. L. acidophilus and L. gasseri, which are prevalent in Western women, were not detected in Korean women. Furthermore, L. rhamnosus, Leuc. lactis, L. coleohominis, and Weissella cibaria, which were not previously reported in the vaginal microbiota of Korean women, were detected. The five most prevalent LABs in vaginal microbiota in Korean women were L. iners, Enterococcus faecalis, L. crispatus, Leuc. lactis, and W. cibaria.

Fabrication of plastic CE (capillary electrophoresis) microchip by hot embossing process (핫 엠보싱 공정을 이용한 플라스틱 CE(capillary electrophoresis) 마이크로 칩의 제작)

  • Cha Nam-Goo;Park Chang-Hwa;Lim Hyun-Woo;Park Jin-Goo
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 2005.06a
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    • pp.1140-1144
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    • 2005
  • A plastic-based CE (capillary electrophoresis) microchip was fabricated by hot embossing process. A Si mold was made by wet etching process and a PMMA wafer was cut off from 1mm thick PMMA sheet. A micro-channel structure on PMMA substrate was produced by hot embossing process using the Si mold and the PMMA wafer. A vacuum assisted thermal bonding procedure was employed to seal an imprinted PMMA wafer and a blank PMMA wafer. The results of microscopic cross sectional images showed dimensions of channels were well preserved during thermal bonding process. In our procedure, the deformation amount of bonding process was below 1%. The entire fabrication process may be very useful for plastic based microchip systems.

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Electrophoretic Analysis of Haemolymph Proteins during Silkworm (Bombyx mori L.) Ontogenesis

  • Staykova, Teodora
    • International Journal of Industrial Entomology and Biomaterials
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    • v.14 no.1
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    • pp.37-44
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    • 2007
  • A study was made of the haemolymph protein spectrum of mulberry silkworm (Bomhyx mori L.) from the first larval instar to imago. Horizontal starch gel electrophoresis was used. Sixteen races and eight F1 interracial hybrids, raised in Bulgaria, were analyzed. During the ontogenesis, a total of 17 protein bands (15 cathodic and 2 anodic) were detected. Distinct dynamics in the haemolymph protein spectrum was observed, in result of different expression during the individual development associated with the processes of growth, histolysis and histogenesis. Based on the ontogenetic dynamics found, a correspondence was assumed between some proteins detected by us using the starch gel electrophoresis and major haemolymph proteins (SP1, SP2, MHPs and Vg) detected by other authors using the polyacrilamide gel electrophoresis. Intraracial and interracial polymorphism was observed in four protein zones. The effect of four polymorphic loci with codominant and null alleles was suggested.

Evaluation of Electrokinetic Flow Mobility Using Isotacho-Electrophoresis Techniques

  • An, J.H.;Joo, Y.H.;Lee, C.Y.;Lee, Y.J.;Park, C.W.
    • Journal of Magnetics
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    • v.16 no.4
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    • pp.444-448
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    • 2011
  • In the present study, we separated the marker particles from the suspending particle mixture solution using isotacho-electrophoresis technique, a novel quantitative ionic particle separation method, in the microchannel. A multiple stacking zone of the suspending particle was visualized with variations in electric field strength, pH value and concentration of the ionic solution. In particular, the electrophoretic mobility of ionic particle (fluorescein) was estimated based on the electrophoretic velocity value measured by the particle image velocimetry. As a result, isotacho-electrophoresis zones were clearly visualized as going downstream in the electric field. The particle migration velocity increased proportional to the applied voltage increase; it was also affected by the pH value variations in the ionic solution.