• 제목/요약/키워드: sodium dodecyl sulfate(SDS)

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A new method for concentration of proteins in the calcareous corpuscles separated from the spargana of Spirometra erinacei

  • PARK Yun-Kyu;PARK Jae-Hwan;GUK Sang-Mee;SHIN Eun-Hee;CHAI Jong-Yil
    • Parasites, Hosts and Diseases
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    • 제43권3호
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    • pp.119-122
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    • 2005
  • Calcareous corpuscles are a characteristic structure found in larval and adult stage cestddes, These corpuscles are known to contain several protein components and to possess protein-binding activity. However, the proteins bound to calcareous corpuscles in situ have not been studied. The present study was undertaken to identify the proteins on calcareous corpuscles. Calcareous corpuscles were purified from the plerocercoids (= spargana) of Spirometra erinacei, and serially dissolved using 0.1 M sulfamic acid solution. Collected supernatants were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining. The results showed that only the fraction remaining after the 19th dissolved fraction contained proteins. A total of 20 protein molecules were detected in gel, with major bands at 56, 53, 46, 40, 35, 29, 28, 24.5, 21, 19, 16, 13, 10 and 8 kDa. In particular, the proteins corresponding to the 21 and 16 kDa bands were most abundant. Our results demonstrated for the first time the protein contents of the calcareous corpuscles of spargana. Further studies on the functions of these proteins are required.

해양미생물 Pseudomonas sp. W7이 생산하는 Extracellular Agarase의 정제 및 Gene Cloning (Purification of Extracellular Agarase from Marine Bacterium (Pseudosmonas sp. W7) and Molecular Cloning of the Agarase Gene)

  • 공재열;배승권
    • KSBB Journal
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    • 제11권1호
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    • pp.37-45
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    • 1996
  • 한국의 남해안에서 한천 분해능이 뛰어난 해양 미 생물을 분리하여 통정한 결과 Pseudomonas 속으로 판명되었으며, 본 연구에서는 이 균을 Halophilic P Pseudomonas sp. W7이라 명영하였다. 이 균주는 호염성 세균으로, 한천의 존재 하에서 높은 효소활성 을 가지는 extracellular agarase를 생산해 내었다. 이 extracellular agarase를 DEAE-Cellulose ani- on exchange chromatography와 gel filtration을 통해 정제하였으며, 정제된 agarase는 SDS-PAGE 를 통해 약 89KDa의 분자량을 지니는 single protein band염을 확인하였다. 한편 agarase의 대량생 산을 위하여 host cell Eo coli JM83과 vector pUC19를 이용하여 gene cloning을 행하였다. Pseudomonas sp. W7의 chromosomal DNA가 삽 입 된 균주 중에 서 agarase activity를 나타내는 E. coli JM83/pSWl과 Eo coli JM83/pSW3를 선멸하 였으며, 전기영통 실험결과 이들은 각각 307Kb, 3.0 K Kb의 chromosomal 단편을 지니고 있음을 확인하였 다. 또한 agarase 유전자가 압입된 변이 균주(B)는 inclusion body 형태의 interacellular agarase를 세포 내에 축적하고 있음이 확인되었다.

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3T3-L1 지방전구세포의 지방분화에서 멜라토닌의 영향 (Effect of Melatonin on Differentiating 3T3-L1 Preadipocytes)

  • 이정근;이영훈;김지현
    • 대한의용생체공학회:의공학회지
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    • 제41권3호
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    • pp.138-145
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    • 2020
  • Adipocytes are the main constituent of adipose tissue. Understanding the molecular basis of adipogenesis is pivotal to finding the therapeutic targets for treatment of obesity. Melatonin is associated with obesity and its mechanism is currently under intensive investigation. The objective of this study was to investigate the effect of melatonin on adipogenesis in differentiating preadipocytes. 3T3-L1 preadipocytes were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 5% calf serum at 37℃ with 5% CO2 in a humidified incubator. Differentiation was induced using DMEM with 10% fetal bovine serum supplemented with MDI two days after cell confluence (day 0). Cells were treated with 0, 10 and 100 μM melatonin on either day 0 or day 5. 72 hours after each treatment, lipid accumulation was measured by oil red O staining. Proteins were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to membranes. As a result, lipid accumulation decreased with melatonin treatment. ERK pathway, activated when differentiation is induced, also decreased with an increase in melatonin concentration. Furthermore, the expression of key adipogenic factors, C/EBPα, C/EBPβ, and PPARγ, were reduced by melatonin treatment. These results imply that melatonin may inhibit the process of adipogenesis and may have a role as a new anti-obesity agent.

인삼엽 단백질의 추출 및 영양가 (Protein Concentrate from Ginseng Leaf and its Nutritive Value)

  • 김지화;이무성;남창우
    • 한국식품과학회지
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    • 제21권3호
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    • pp.441-445
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    • 1989
  • 인삼엽 단백질의 식량화를 위하여 엽단백질 농축물(LPC)을 조제하여 그 아미노산 조성과 인공소화율을 검토하였다. 인삼엽을 $-20^{\circ}C$ 냉아세톤으로 처리하고 0.2% NaOH(0.5% SDS, 0.5% 2-ME 함유)로 단백질을 추출하며 인삼 LPC를 얻었다. 인삼 LPC의 일반성분으로는 지방과 회분이 1% 미만이었고 단백질은 75% 내외로 많았으며 총 당은 약 5% 이었다. 총 사포닌 함량은 약 1.2%이고 panaxadiol과 panaxatriol의 비는 1.67이었다. 아미노산 조성은 aspartic acid의 함량이 약 13.4%로 가장 많았으며 methionine의 함량은 매우 낮았다. 필수아미노산토 고루 들어 있었고 총 아미노산에 대한 필수아미노산은 54.5%이였고 E/T ratio는 3.02이었다. LPC의 제 1제한아미노산은 함황아미노산이고, 그 amino acid score는 43.1이었다. Pepsin및 trypsin을 사용한 인삼 LPC의 소화율은 비교단백질인 milk casein에 비하며 낮았다.

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초기 임신돈에 있어서 자궁분비단백질의 전기영동상과 면역억제력에 관한 연구 (A Study on Electrophoretic Pattern and Immunosuppressive Activity of Uterine Secretory Protein in Early Pregnant Sows)

  • 김경순;정영채;김창근;한기영;김형태;윤종택;김종대;최선호
    • 한국가축번식학회지
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    • 제13권3호
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    • pp.179-187
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    • 1989
  • The experiment was carried out to study the profile of uterine specific protein during early pregnancy in sows and to test it's immunosuppressive activity. Uterine protein samples were obtained by flushing the uterine horn on Day 3, 6, 9, 12 and 15 of the estrous cycle and the pregnancy respectively and the protein concentration of each sample was determined. The change of uterine protein was examined by sodium dodecyl sulfate(SDS)-PAGE. The immunosuppressive activity of uterine secretory protein was investigated according to the lymphocyte blastogenesis response to mitogen. The results of this experiment are summarized as follows ; 1. The uterine protein during estrous cycle and early pregnancy was relatively constant up to Day 9, but increased on Day 12. Maxium total protein values were found on Day 15. The concentration of serum proteins were about 82-95 mg during estrous cycle, but decreased to about 70-82 mg during early pregnancy. 2. The proteins components similar electrophoretic patterns(PAGE) that were no differences (band ; a, b, c, d, e, f, g, I) on Days 3, 6 and 9 of the estrous cycle and pregnancy. But there were more 2 bands specifically on Day 12 of the pregnancy and on Day 15 of estrous cycle and showed more 4 bands on Day 15 of early pregnancy. They seemed to be acidoprotein and their average molecular weight were 38,000, 22,300 and 12,600. 3. When uterine protein were added 500$\mu\textrm{g}$/ml, there was no immunosuppresive activity on Day 3 of estrous cycle and lymphocyte blastogenesis was slightly suppressed on Day 3 of pregnancy. The immunosuppressive activity on Day 9 of estrous cycle and pregnancy appeared in 500$\mu\textrm{g}$/ml and 150$\mu\textrm{g}$/ml, respectively the uterine protein on Day 12 and 15 showed immunosuppresive activity, which at the level of 150$\mu\textrm{g}$/ml during non-pregnancy and at the level of 100 to 125$\mu\textrm{g}$/ml during early pregnancy, respectively.

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암의 복수액에 존재하는 지방분해 및 식욕감퇴 인자에 미치는 고려인삼의 영향 (EFFECT OF RED GINSENG POWDER ON LIPOLYTIC AND ANOREXIGENIC FACTOR (TOXOHORMONE-L) FROM CANCEROUS ASCITES FLUID)

  • Okuda Hiromichi;Masuno Hiroshi;Lee Song Jae
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1984년도 학술대회지
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    • pp.145-152
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    • 1984
  • 여러 종류의 종양 환자에서 점차적인 체중 감소가 일어나는 과정에서, 체내에 저장된 지방이 고갈됨이 관찰되었다. 종양이 커가는 동안의 체내 지방의 고갈은 혈장의 유리지방산의 증가와 연관이 있다. 최근에, 우리는 간암, 또는 난소암 환자의 복수액과 악성 임파종 환자의 늑막액은(in vitro에서) 흰쥐의 지방조직에 유리지방산의 방출을 촉진시킴을 발견하였다. Toxoh-ormone-L(독성 호르몬-L)이라고 명명한 이 지방분해 인자를 간암 환자의 복수액에서 분리, 정제하였다. 이 분리된 물질은 disc gel electrophoresis와 ${\btea}$-m-ercaptoethanol 존재하의 sodium dodecyl sulfate(SD5)-acrylamide gel electrophoresis상에서 단일띠를 나타내었다. 이것의 분자량은 각각 SDS-acrylamide gel electrophoresis와 analytical 초원심분리에 의하여 $70,000{\~}75,000$과 65,200의 분량을 가짐이 확인되었다. 흰쥐에 Toxohormone-L의 주사는 사료와 물의 섭취를 현저히 억제하였으며, 이 Toxohormone-L의 주사 5시간후에 이러한 억제효과가 나타났다. Toxohormone-L의 지방분해의 작용을 억제하는 억제물질이 고려인삼 뿌리의 물 추출물에 존재함이 입증되었다. 즉, 인삼 분말을 5배량의 증류수에 현탁시켜 추출하고 감압 농축한 다음 증류수에 투석시켰다. 투석막안의 지방 성분을 석유 에테르로 제거하고, 사포닌은 수포화된 부탄올로 제거하였다. 이렇게 제거하고 남은 여액을 활성탄으로 처리하고 DEAE-cellulose column에 통과시켰다. 이러한 과정으로 유효한 분획 (인삼분획 3)을 얻었으며, 보다 더 순수한 정제가 지금 진행중에 있다. 이러한 실험 결과를 바탕으로 볼 때, 암환자에 고려인삼 분말의 투여는 암조직에서 분비되는 Toxohormone-L(독성 호르몬-L)의 작용을 억제함으로서 암환자의 체중 감소를 방지하고, 식욕감퇴를 개선할것으로 생각된다.

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Effect of Au-ionic Doping Treatment on SWNT Flexible Transparent Conducting Films

  • 민형섭;정명선;최원국;김상식;이전국
    • 한국재료학회:학술대회논문집
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    • 한국재료학회 2012년도 춘계학술발표대회
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    • pp.111.1-111.1
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    • 2012
  • Interest in flexible transparent conducting films (TCFs) has been growing recently mainly due to the demand for electrodes incorporated in flexible or wearable displays in the future. Indium tin oxide (ITO) thin films, which have been traditionally used as the TCFs, have a serious obstacle in TCFs applications. SWNTs are the most appropriate materials for conductive films for displays due to their excellent high mechanical strength and electrical conductivity. In this work, the fabrication by the spraying process of transparent SWNT films and reduction of its sheet resistance on PET substrates is researched Arc-discharge SWNTs were dispersed in deionized water by adding sodium dodecyl sulfate (SDS) as surfactant and sonicated, followed by the centrifugation. The dispersed SWNT was spray-coated on PET substrate and dried on a hotplate. When the spray process was terminated, the TCF was immersed into deionized water to remove the surfactant and then it was dried on hotplate. The TCF film was then was doped with Au-ionic doping treatment, rinsed with deionized water and dried. The surface morphology of TCF was characterized by field emission scanning electron microscopy. The sheet resistance and optical transmission properties of the TCF were measured with a four-point probe method and a UV-visible spectrometry, respectively. This was confirmed and discussed on the XPS and UPS studies. We show that 87 ${\Omega}/{\Box}$ sheet resistances with 81% transmittance at the wavelength of 550nm. The changes in electrical and optical conductivity of SWNT film before and after Au-ionic doping treatments were discussed. The effect of Au-ion treatment on the electronic structure change of SWNT films was investigated by Raman and XPS.

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Ink-jet 프린팅용 CoAl2O4 고화도 나노 무기 잉크 제조 및 프린팅 특성평가 (Synthesis and Characterization of CoAl2O4 Glazed Blue Ceramic Ink for Ink-Jet Printing)

  • 이기찬;윤종원;김진호;황광택;한규성
    • 한국재료학회지
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    • 제24권2호
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    • pp.73-80
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    • 2014
  • Ink-jet printing technology has been widely attractive due to its facility for direct and fine printing on various substrates. Recent studies have focused on expanding the application of ink-jet printing technology from general consumer use and design companies to the prototype production of precision parts and parts manufacturing. The use of ink-jet printing technology in decorated tableware, tiles, and other ceramic products also has many advantages. The printing process is fast and can be adaptable to various kinds of objects because there is no direct contact point between the printer and the substrates to be printed. For application to ceramic product decoration, inks containing highly dispersed inorganic nano-pigments are required. Here we report the synthesis and characterization of blue $CoAl_2O_4$ nanopigment for ink-jet printing. Blue ceramic ink based on the obtained $CoAl_2O_4$ pigment was prepared by dissolving $CoAl_2O_4$ pigment in a mixed solution of ethylene glycol and ethanol with volume ratios of 7:3 and 8:2, respectively, to obtain the appropriate viscosity for ink-jet printing. The ink solution contained 15 wt% of $CoAl_2O_4$ pigment and Cetyltrimethyl ammonium bromide(CTAB) and Sodium dodecyl sulfate(SDS) as dispersive agents. The prepared blue ceramic ink was stably jetted and formed a sphere-shaped droplet from an ink-jet printer.

경주 적혈구막 단백의 전기영동법에 의한 분석 -빠른 적혈구 침강 속도와의 관계- (Electrophoretic analysis of the major proteins of race horse erythrocyte membrane: Their relation to fast erythrocyte sedimentation rate)

  • 박영우;이시영
    • 대한수의학회지
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    • 제31권3호
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    • pp.259-264
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    • 1991
  • The proteins of the race horse erythrocyte membrane were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate(SDS-PAGE), and their relations to the fast erythrocyte sedimentation rate(ESR) of the race horse were investigated. The erythrocyte sedimentation rate of race horse were very fast compared with the human one(33 times <$90^{\circ}-plastic-ESR/30m$> and 25 times <$90^{\circ}-micro-ESR/30m$> as fast as the human one) are reported previously. Although the general protein profiles of the race horse erythrocyte membranes were almost similar to that of human, band 3 content was showing higher in race horse (34.7%) than in human (25.3%). The glycoprotein profiles of the race horse erythrocyte membranes revealed by periodic acid Schiff's(PAS) stain showed a marked difference from that of human. The PAS-1(glycophorin) and PAS-2(sialoglycoprotein) present in human erythrocyte memo brane were almost absent from the Holstein and race horse erythrocyte membranes, but PAS-2 was more in only race horse from that of human. Instead, the bovine erythrocyte membranes showed a strong PAS-B near the origin of the electrophorograms and the race horse erythrocyte membranes showed a strong PAS-negative band near the end of the electrophorograms, which is named as PAS-E in this study. These results suggest that the fast sedimentation rate of race horse erythrocyte is due in part to the presence of more band 3 protein fraction and PAS-E glycoproteins in the race horse erythrocytes.

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Neutralization of Human Papillomavirus by Specific Nanobodies Against Major Capsid Protein L1

  • Minaeian, Sara;Rahbarizadeh, Fatemeh;Zarkesh-Esfahani, Sayyed Hamid;Ahmadvand, Davoud;Broom, Oliver Jay
    • Journal of Microbiology and Biotechnology
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    • 제22권5호
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    • pp.721-728
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    • 2012
  • The human papillomavirus (HPV) is the main cause of cervical cancer in developing countries. Rapid diagnosis and initiation of treatment of the HPV infection are critical. Various methods have been employed to reduce the immunogenicity of antibodies targeting HPV serotypes. Nanobodies are the smallest fragments of naturally occurring single-domain antibodies with their antigen-binding site compromised into a single domain. Nanobodies have remarkable properties such as high stability, solubility, and high homology to the human VH3 domain. In this study, a phagemid library was employed to enrich for nanobodies against the L1 protein of the human papilloma virus. Binding reactivity of the selected clones was evaluated using phage enzyme-linked immunosorbent assay (phage-ELISA). Finally, two nanobodies (sm5 and sm8) with the best reactivity against the Gardasil vaccine and the purified HPV-16 L1 protein were expressed and purified using a $Ni^+$-NTA column. The accuracy of expression and purification of the nanobodies was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting assays. In vitro studies demonstrated that neutralization was achieved by the selected nanobodies. The ease of generation and unique features of these molecules make nanobodies promising molecules for the new generation of HPV diagnosis and therapy.