• Title/Summary/Keyword: Gel fraction

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Proteomic Analysis of the Hydrophobic Fraction of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood

  • Jeong, Ju Ah;Lee, Yoon;Lee, Woobok;Jung, Sangwon;Lee, Dong-Seong;Jeong, Namcheol;Lee, Hyun Soo;Bae, Yongsoo;Jeon, Choon-Ju;Kim, Hoeon
    • Molecules and Cells
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    • v.22 no.1
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    • pp.36-43
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    • 2006
  • Mesenchymal stem cells (MSCs) are promising candidates for cell therapy and tissue engineering, but their application has been impeded by lack of knowledge of their core biological properties. In order to identify MSC-specific proteins, the hydrophobic protein fraction was individually prepared from two different umbilical cord blood (UCB)-derived MSC populations; these were then subjected to two-dimensional (2D) gel electrophoresis and peptide mass fingerprinting matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF)-mass spectrometry (MS). Although the 2D gel patterns differed somewhat between the two samples, computer-assisted image analysis identified shared protein spots. 35 spots were reliably identified corresponding to 32 different proteins, many of which were chaperones. Based on their primary sub-cellular locations the proteins could be grouped into 6 categories: extracellular, cell surface, endoplasmic reticular, mitochondrial, cytoplasmic and cytoskeletal proteins. This map of the water-insoluble proteome may provide valuable insights into the biology of the cell surface and other compartments of human MSCs.

The Effect of Temperature and Exposure Time on UV-curing of Urethane Acrylate (UV 챔버 내부 온도 및 체류시간이 Urethane Acrylate의 경화속도에 미치는 영향)

  • Moon, Seung-Ho;Kim, Kwang-Hee;Kim, Jeong-Sik
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.13 no.9
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    • pp.3822-3826
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    • 2012
  • We have studied the effect of temperature and exposure time on the UV curing rate of Urethane Acrylate. UV dose has been modulated by changing the electric power, and temperature has been controled by coating rate and water temperature of the coating roll. Curing rate was evaluated by measuring gel fraction of the cured film. It has been found that effect of coating velocity on the gel fraction of the final film is predominant, but role of temperature of the UV chamber is negligible. These observations imply that exposure time of UV light is critical to have a fully cured film. To better understand the effect of design and operating condition of UV coater on the behavior of UV curing of Urethane Acrylate, we have also conducted heat-flux analysis of UV chamber.

Tumour Specific and Immunosuppressive Components in Soluble Cell Extracts from Ovine Squamous Cell Carcinoma (면양(緬羊) Squamous Cell Carcinoma의 세포추출액(細胞抽出液)중에 함유된 종양특이(腫瘍特異) 및 면역기능저하물질(免疫機能低下物質))

  • Jun, Moo Hyung
    • Korean Journal of Agricultural Science
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    • v.13 no.2
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    • pp.299-310
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    • 1986
  • The ovine squamous cell carcinoma (OSCC)-specific and immunosuppressive properties of OSCC extracts were investigated by using the techniques of lymphocyte blastogenicity, acid dissociation-ultrafiltration and gradient polyacrylamide gel electrophoresis. It was found that OSCC extracts contained two major and one minor protein peaks by Sephadex gel fractionation. Two major peaks bear substantial amount of immunoglobulins, antigen-antibody complex and OSCC-specific fractions, and the minor peak includes immunosuppressive materials. OSCC-specific components were detected at the molecular weights of 10,000 to 100,000 daltons in the major peaks and immunosuppressive materials at the fractions with the molecular weight of 10,000 to 100,000 and < 10,000 daltons in the minor peak. When the fractions were further separated by gradient polyacrylamide gel electrophoresis, the OSCC-specific antigens were found in the slice number 4 to 6 in fraction III, and immunosuppressive materials, in the slice numbers 9 to II in fraction V. The present results were considered to provide a basis for preparation and purification of OSCC-specific and immunosuppressive materials from the crude OSCC extracts.

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Isolation of Angiotensin Converting Enzyme Inhibitor from Doenjang (전통된장으로부터 Angiotensin Converting Enzyme 저해물질의 분리)

  • Kim, Seung-Ho;Lee, Yun-Jin;Kwon, Dae-Young
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.848-854
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    • 1999
  • Inhibitory compounds of angiotensin converting enzyme (ACE) were separated from Doenjang (traditional Korean fermented soybean paste). Water extracts from Doenjang which showed ACE inhibitory activity were separated with gel permeation chromatography (GPC), in which two fractions with high ACE inhibitory activities were obtained. The first fraction from GPC was further isolated by semi-preparative reverse phase preparative-HPLC (high performance liquid chromatography) and 2-dimensional electrophoresis/thin layer chromatography (TLC). The purified spot had molecular weight of 759 daltons and ninhydrin-positive non-peptide. The second fraction from GPC was also further isolated by semi-preparative reverse phase HPLC and $NH_2-column$ HPLC. One fraction with high ACE inhibitory activity was purified and characterized. Molecular weight of this fraction by LC-MS was 272.34 daltons. The active fraction was identified as Arg-Pro with ACE $IC_{50}$ of $92\;{\mu}M$.

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Studies on the Changes of Taste Compounds during Soy Paste Fermentation (III) (된장 숙성 중 정미 성분의 변화에 관한 연구(III))

  • 김미정;이혜수
    • Korean journal of food and cookery science
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    • v.9 no.4
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    • pp.261-265
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    • 1993
  • For the purpose of supplying the information to improve the acceptability of soy paste as the condi-ment, the changes of peptide were determined. The results were as follows; Average peptide length were decreased. It was 102 at 0 day, 15 at 10 day and 4.1 at 180 day. Peptide fraction were the same as in 60 day and 180 day. Low molecular weight peptide were not changed greatly during fermention. Peptide identified in 180 day fermentation were Ala-Ser, Gly-Glu, Glu-Ser, Asp-Glu, Asp- Tyr, Asp-Ala-Ser, Ala-Ser-Glu, Glu-Ser-Ala, and Ala-Lys-Met. In the characteristics of bitter peptide in 180 day fermentation, soy paste itself didn't show bitter taste', solvent extration fraction I'showed bitter taste. After gel chromatography, fraction I, fraction II and fraction III were obtained and fraction II were bitter peptide of low molecular weight. After gel chromatography', solvent extration fraction 2'(water extration) were divided into fraction IV, V, VI,VII and VIII. Fraction IV, V and VI showed bitter taste. Amino acids composition of the fractions showing bitter taste were like that; fr. 1: Glu- (Asp, Pro, Val, lie or Leu)-Met fr. II Pro-(Glu, Val, Phe)-lle or Leu fr. IV: Glu-(Asp, Ala, Tyr, Leu of lie)-Phe fr. V: Ala-(Met, Glu, Pro)-lle or Leu fr. VI: Asp-(Phe, Ser, fly)-Val.

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Characteristic Component of Rehmanniae Radix Preparata Compared to Rehmanniae Radix and Rehmanniae Radix Crudus (숙지황, 건지황 및 생지황 중 숙지황의 특이성분 검색)

  • Hong, Sun Pyo;Kim, Young Chul;Kim, Kyeong Ho;Park, Jeong Hill;Park, Man Ki
    • Analytical Science and Technology
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    • v.6 no.4
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    • pp.401-404
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    • 1993
  • Rehmanniae Radix Preparata is manufactured with Rehmanniae Radix accoding to KP V. For quality control of Rehmanniae Radix Preparata, its standard component is required. The methanol extracts of Rehmanniae Radix crudus, Rehmanniae Radix, Rehmanniae Radix preparata were divided into the three groups of ether, butanol and aqueous fraction by liquid-liquid separation. In the comparative TLC of ether fraction, the characteristic component of Rehmanniae Radix preparata was found. The ether fraction was evaporated and separated on the silica gel column with chloroform-methanol and further separated on the preparative silica gel TLC with chloroform-methanol-water. The component was illucidated as 5-(hydroxymethyl)-2-furancarboxaldehyde(5-HMF). 5-HMF was not found in Rehmanniae Radix crudus and found in Rehmanniae radix in much less Quantities than Rehmanniae Radix Preparata.

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Antioxidant Activity of Amino Acid-Xylose Browning Reaction Products 3. Isolation of Antioxygenic Substances from Browning Reaction Products by Solvent Extraction, Column Chromatography and Gel Filteration (Amino산-Xylose 갈변반응물질의 항산화성 3. 용매추출, Column Chromatography와 Gel여과에 의한 항산화성 갈변물질의 분리)

  • YOU Byeong-Jin;LEE Kang-Ho;LEE Jong-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.4
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    • pp.273-281
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    • 1987
  • In this work the antioxidant effects of browning reaction products prepared by xylose-tryrtophan reaction system were discussed. The antioxygenic brown pigments were separated by solvent extraction, and column chromatography and isolated by gel filteration. The functional groups of the brown pigments which had antioxidant activity were examined. The brown pigments extracted with methanol showed antioxidant effect and were fractionated in 5portions on DEAE-cellulose column. The elutes with methanol: acetic acid(10:30 v/v sol n(A), methanol: chloroform(95:5 v/v) sol n(C), and chloroform: acetic acid(10:30 v/v) sol n(E) only showed antioxidant activity and their compositions were 22.43, 21.51 and $34.43\%$ respectively. When each fraction on DEAE-cellulose column was reseparated on Sephadex LH-20 column, 2 fractions were obtained from portion A and C respectively. Molecular weights of A, C and E fraction of brown pigments were from 2,600 to 3,700. By elucidation of IR spectra, the pigment fractions which showed a strong antioxidant activity were tearing the indole group. It is suggested that the antioxidant function of the brown pigment is due to hydroxy and amino group. A higher activity of the brown pigment fraction E might be attributed to carboxylic acid or carboxylic ester compounds.

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Identification and Isolation of Juvenile Hormone Binding Protein from Hemolyrnph of Lymantria dispar L. (매미나방(Lymantria dispar)에서 Juvenile Hormone Binding Protein(JHBP)의 확인 및 정체)

  • 이인희;김학열
    • The Korean Journal of Zoology
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    • v.34 no.2
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    • pp.196-202
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    • 1991
  • Juvenile honnone binding protein (JHBP) was identified in the last instar larval hemolymph of Lymantria dispar using dextran coated charcoal (DCC) binding assay and gel filtration. The p1 value of JHBP was estimated to be 5.3. JHBP was partially pudfied by polyethylene glycol(PEG) precipitation, DEAE-cellulose ion-exchange chromatography and gel filtration, and was confirmed by DCC binding assay.

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Purification of Isocitrate lyase Produced from Microbacterium laevaniformans (Microbacterium laevaniformans가 생성하는 Isocitrate lyase의 정제)

  • 서승교;김정호
    • Journal of Environmental Science International
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    • v.7 no.6
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    • pp.853-857
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    • 1998
  • Purification of the isocitrate lyase extracted from Microbacterium laevaniformans was investigated. The isocitrate lyase was purified 43.6 folds by the following continuous treatment with ammonium sulfate fraction, DEAE-cellulose, DEAE-sephacel and Sephadex G-200 chromatography. The purified isocitrate lyase was showed to be a single protein band by polyacrylamide gel electrophoresis. The molecular weight of the purified isocitrate lyase was estimated 54,000 Da by the SDS-polyacrylamide gel electrophoresis. The Km and Vmax values for isocitrate were estimated to be 0.83mM and 0.33units/ml, respectively. Activity of isocitrate lyase was inhibited by cystein-HCl and glutathione.

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Inhibitory Effect of Artemisia princeps Pampan.. Extract on Growth of Cancer Cell Lines (쑥(Artemisia princeps Pampan) 추출 성분의 암세포증식 억제효과)

  • 황윤경
    • Journal of Nutrition and Health
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    • v.31 no.4
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    • pp.799-808
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    • 1998
  • To investigate the antitumor activity of mugwort (Artemisia princeps Pampan), petroleum ether extract of mugwork was partially purified by a silica gel chromatography. Among several fractions, the fraction which was obtained under the elution with acetone, showed potent cytotoxicity against mouse leukemia cell line(Ll210), human colon cancer cell line (HCT-48) and human hepatoma cell line (Hep G2) , but was less effective with normal cell line(mouse embryo cell). Acetone fraction appeared to be glycolipid by Benedict test and the major fatty acids of the lipid were C16 ; 0 , C 18: 3by GC/MS analysis.

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