• Title, Summary, Keyword: SDS-PAGE

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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.

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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Identification of Lactic Acid Bacteria in Kimchi Using SDS-PAGE Profiles of Whole Cell Proteins

  • Kim, Tae-Woon;Jung, Sang-Hoon;Lee, Ji-Yeon;Choi, Sun-Kyu;SUN-HEE-PARK;JAE-SUN-JO
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.119-124
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    • 2003
  • This study was conducted to evaluate the practical usefulness of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PACE) fingerprinting of whole cell proteins far the identification of lactic acid bacteria in Kimchi. SDS- PACE of whole cell proteins of the reference strains and lactic acid bacteria isolated from Kimchi yielded differential banding patterns that were highly specific fingerprints, thus making it possible to identify. Identification of the isolates from Kimchi was achieved by comparing the SDS-PAGE fingerprints of isolates to those of reference strains. In addition, the reliability of SDS-PAGE was examined by comparing the results with those of the APL 50 CHL system assay and 16S rRNA gene sequence. SDS-PACE assay showed a different identity to reference strains, while the APL 50 CHL system and 16S rRNA gene sequence could not distinguish a few strains. Therefore, SDS-PAGE of the whole cell proteins is a specific and a reliable method that will be useful for the identification of lactic acid bacteria in Kimchi to the species level, and can be used as an alternative or complementary identification method.

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.

Distribution of Dominant Bifidobacteria in the Intestinal Microflora of Korean Adults and Seniors, Identified by SDS-PAGE of Whole Cell Proteins and 16S rDNA Sequence Analysis

  • KIM TAE WOON;SONG HEE SUNG;KIM HAE YEONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.388-394
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    • 2005
  • In order to investigate the distribution of dominant Bifidobacterium species in intestinal microflora of Korean adults and seniors, SDS-PAGE profiles of whole cell proteins were used for the identification of bifidobacteria. To confirm the reliability of SDS-PAGE, the Bifidobacterium species identified by SDS-PAGE of whole cell proteins were validated by using 16S rDNA sequencing analysis. The results of SDS­PAGE corresponded well with those determined by the analysis of 16S rDNA sequencing. Based on the analysis of SDS-PAGE patterns on unidentified fecal strains which showed positive in fructose-6-phosphate phosphoketolase activity, B. adolescentis, B. longum, and B. bifidum were identified in the feces of adults, and B. adolescentis, B. longum, B. bifidum, B. breve, and B. dentium were identified in those of seniors. In most of the fecal samples tested, the predominant Bifidobacterium species consisted of only a few species, and differences in the distribution and numbers of Bifidobacterium species were observed between adults and seniors. B. adolescentis and B. longum were found to be the most common species in feces of adults, but not in seniors. Accordingly, the distribution and abundance of bifidobacteria in the human intestinal microflora varied depending on the age of hosts.

Change in Soluble and Insoluble Proteins during Embryonic Development of the Silkworm, Bombyx mori (누에알의 배자발육에 따른 가용성 및 불용성 단백질의 변화)

  • 성주일
    • Journal of Sericultural and Entomological Science
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    • v.33 no.1
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    • pp.1-5
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    • 1991
  • PBS soluble- and insoluble-extracts form silkworm eggs were analysed by native-PAGE and SDS-PAGE. During the embroyonic development, soluble proteins which are mainly composed of ESP, vitellin and 30K proteins showed similar degradation pattern in both electrophoretic analyses. Several peptides which seemed to be intermediated forms of yolk proteins were detected in latter part of embryogenesis, while a protein band newly appeared in one day elapsed after acid treatment. When insoluble extracts from silkworm eggs were analysed with SDS-PAGE, several peptides were detected at the later stages of the embryonic development, and newly hatched larvae. These peptides are considered to be structure proteins for embryonic morphogenesis.

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Effect of ${\gamma}-irradiation$ on the Physicochemical Properties of Soy Protein Isolate and Whey Protein Concentrate (감마선 조사가 Soy Protein Isolate와 Whey Protein Concentrate의 이화학적 성질에 미치는 영향)

  • Cho, Yong-Sik;Song, Kyung-Bin
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1488-1494
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    • 1999
  • Effect of ${\gamma}-irradiation$ on the SDS-PAGE pattern, secondary structure content, the solubility of commercial soy protein isolate (SPI) and whey protein concentrate (WPC) was investigated. The change in the subunit molecular weight of SPI and WPC irradiated in aqueous solution or dried state was studied using SDS-PAGE. The SDS-PAGE pattern of SPI irradiated in aqueous solution revealed the fragmentation and aggregation of the subunit protein. For WPC irradiated in aqueous solution. fragmentation of the subunit protein up to 10 kGy was observed. In contrast, ${\gamma}-irradiation$ of SPI and WPC in dried state did not cause any significant changes in the SDS-PAGE pattern. The change In the secondary structure of irradiated SPI and WPC solution was studied using circular dichroism. The aperiodic structure content of SPI and WPC solution increased at higher irradiation doses, which suggests that ${\gamma}-irradiation$ caused the disruption of the ordered structure of SPI and WPC solution. Gamma-irradiation also caused the change in solubility of SPI and WPC in dried state.

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Changes of SDS-PAGE Pattern and Allergenicity of BSA and BGG in Beef Extract Treated with Heat and High Pressure (물리적 처리에 따른 우육추출물중의 BSA와 BGG단백질의 SDS-PAGE패턴 및 항원성의 변화)

  • Han, Gi-Dong;Fan, Jiang Ping;Suzuki, Atsushi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.5
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    • pp.594-599
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    • 2006
  • In our previous report, we indicated that not only BSA but also BGG played an important role in the allergenicity of beef. In this study, the effect of heat or high-pressure treatments to beef extract on the SDS-PAGE patterns was examined. The antigenicity of each treated samples was also investigated by Western blots assay with the sera of BGG-positive beef allergic patients. The BGG band and its antigenicity slightly disappeared but not generally in $100^{\circ}C$ group, indicating $100^{\circ}C$ treatment is not sufficient to totally eliminate the antigenicity of beef allergens. Compared with BGG band, BSA band significantly disappeared in SDS-PAGE with $100^{\circ}C$ treatment, indicating BSA is more heat- sensitive than BGG. When the beef extract was heated at $120^{\circ}C$, not only BSA but also BGG bands was largely disappeared in both SDS-PAGE and Western blots. High pressure (HP) treatment even at 600 MPa did not affect SDS-PAGE and Western blots pattern of BSA. On the contrary, BGG treated with HP showed visible changes in SDS-PAGE. 600 MPa treatment significantly reduced the antigencity. Interestingly, these behaviors of BGG were not found in the same experiments with pure BGG treated with HP. From these results, it was speculated that some kinds of proteolytic enzymes in beef extracts were involved in the BGG molecular degradation by HP treatment. The aging experiments of beef extracts treated with HP supported this hypothesis. Further studies are needed to clarify the function and working mechanism of enzymes associated with BGG degradation in beef extracts by HP treatment.

Proteins and Amino Acid Composition of Korea Ginseng Classified by Years (한국인삼의 연근별 단백질 및 아미노산 조성)

  • Choi, Cheong;Yoon, Sang-Hong;Bae, Man-Jong;An, Bong-Jean
    • Korean Journal of Food Science and Technology
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    • v.17 no.1
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    • pp.1-4
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    • 1985
  • For the systematic investigation of biochemical characteristics of Korean ginseng protein by years, protein fractions were analyzed by the techniques of polyacrylamide gel electrophoresis, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration, while the amino acid composition was studied by amino acid autoanalyzer. Results of polyacrylamide gel electrophoresis and SDS-PAGE showed a few difference in pattern and number of bands depending on the age of the root. However, the number of bands obtained from polyacrylamide gel electrophoresis and SDS-PAGE was 8 and 7 to 11, respectively. When water extracted proteins were fractionated by Sephadex G-200, the main peak among 2 peaks was collected and lyophillized. Its mol. wt. was extimated to be 43,000 by the SDS-PAGE method. In amino acid composition of water extracted protein and main fraction of gel filtration, arginine content was the highest, 47.17% in water extracted protein and 57.36% in main fraction followed by glutamic acid and asparatic acid. On the contrary, cystine and methionine contents were very low in both cases.

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Characteristics of IEF Patterns and SDS-PAGE Results of Korean EPO Biosimilars

  • Kang, Min-Jung;Shin, Sang-Mi;Yoo, Hey-Hyun;Kwon, Oh-Seung;Jin, Chang-Bae
    • Bulletin of the Korean Chemical Society
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    • v.31 no.9
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    • pp.2493-2496
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    • 2010
  • Erythropoietin (EPO) is mainly produced in kidney and stimulates erythropoiesis. The use of recombinant EPOs for doping is prohibited because of its performance enhancing effect. This study investigated whether biosimilar EPOs could be differentiated from endogenous one by iso-electro-focusing plus double blotting and SDS-PAGE for antidoping analysis. The established method was validated with positive control urine. The band patterns were reproducible and meet the criteria, which was made by world anti doping agency (WADA). Isoelectric focusing was conducted in pH range 2 to 6. Recormon (La Roche), Aropotin (Kunwha), Epokine (CJ Pharm Co.), Eporon (Dong-A), Espogen (LG Life Sciences), and Dynepo (Shire Pharmaceuticals) were detected in basic region. All biosimilars showed discriminative isoelectric profiles from endogenous EPO profiles, but they showed different band patterns with the reference one except Epokine (CJ Pharm Co.). Next, SDS-PAGE of biosimilar EPOs resulted in different molecular weight patterns which were distributed higher than endogenous EPO. Commercial immune assay kit as an immune affinity purification tool and immobilized antibody coated magnetic bead were tested for the purification and concentration of EPO from urinary matrix. The antibody-coated magnetic bead gave better purification yield. The IEF plus double blotting and SDS-PAGE with immunoaffinity purification method established can be used to discriminate biosimilar EPOs from endogenous EPO.