• Title/Summary/Keyword: Western Blot

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Production and Characterization of Monoclonal Antibodies to Escherichia coli (ATCC 8739)

  • Yoo, Dohng-Hyun;Lee, Young-Ha;Jung, Jae-Deuk
    • Journal of Microbiology and Biotechnology
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    • v.5 no.6
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    • pp.353-358
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    • 1995
  • Escherichia coli causes intestinal and extraintestinal infections and has been an indicator of fecal pollution in water and food. BALB/c mouse was immunized by injection of somatic E. coli (ATCC 8739) cells to produce monoclonal antibodies. Splenocytes of mouse were fused with myeloma cells (Sp2/0-Ag14). Two hybridomas secreting monoclonal antibodies were established after being cloned. In SDS-PAGE analysis of E. coli antigens 37 protein profiles appeared from 14 kDa to 182 kDa. Western blot analysis using polyclonal antibodies demonstrated that protein antigens of 41 kDa, 38.2 kDa and 31.7 kDa were immunodominant. Monoclonal antibodies DY-CM1 and DY-CM2 recognized 31.7 kDa and 2.0 kDa antigens in Western blot analysis, respectely.

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First Report of Sweet potato latent virus and Sweet potato chlorotic stunt virus Isolated from Sweet Potato in Korea

  • Yun, W.S.;Lee, Y.H.;Kim, K.H.
    • The Plant Pathology Journal
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    • v.18 no.3
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    • pp.126-129
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    • 2002
  • Infected sweet potato (Ipomoea batatas) showing symptoms of sunken veins, stunting, mosaic, and mottling were collected from Gimje, Cochang, Iksan, and Haenam provinces in Korea. Electron microscopic (EM) observation of the infected tissue revealed rod and filamentous rod type virus particles of various lengths. Western blot analysis of the protein samples extracted from infected sweet potato and partially purified virus identified the isolates as Sweet potato feathery motile virus (SPFMV), Sweet potato latent virus (SwPLV), and Sweet potato chlorotic stint virus (SPCSV). Sweet potatoes were occasionally infected with more than one of these viruses. This is the first report of SwPLV and SPCSV in Korea.

Induction of Porcine Oocytes Maturation by MAP Kinase (MAP Kinase에 의한 돼지 난성숙의 유기)

  • 장규태;박미령;윤창현
    • Korean Journal of Animal Reproduction
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    • v.22 no.3
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    • pp.277-286
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    • 1998
  • The effect of MAP kinase activity on maturation of porcine oocytes was investigated. MAP kinase was detected by immunofluorescence staining in nucleus of oocytes just before entering GVBD (germinal vesicle breakdown)stage. In a Western blot with GV (germinal vesicle) from these oocytes (cultured for 25 hours), a shift of MAP kinase band a, pp.ared, suggesting an activated stage of the kinase. No activity was shown in the blot with GV isolated from, oocytes cultured for 0 hour. To confirm that activation of MAP kinase induce GVBD, we microinjected MAP kinase purified from matured oocytes starfish into the cytoplasm of oocytes in GV stage (cultured for 0 hour). The injected MAP kinase did not cause early a, pp.arance of GVBD. No oocytes showed GVBD state until 20 hours of culture. Activity of MAP kinase did not increase significantly after the injection. When the exogenous MAP kinase was injected into GV, GVBD was induced in about 20% of oocytes cultured for 5 to 10 hours. These results suggest that MAP kinase is traslocated to nucleus and function as a factor inducing GVBD.

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The effcect of Gorynju and a Soju extract Psoraleae fructus on Melanin synthesis of B16 melanoma cells (보골지(補骨脂)의 고량주(高梁酒), 소주(燒酒) 추출물이 B16 melanoma 세포주의 멜라닌 합성에 미치는 영향)

  • Seo, Hyeong-Sik
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.21 no.1
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    • pp.96-103
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    • 2008
  • Objective : This research was carried out to compare the effect of Gorynju and a Soju extract Psoraleae fructus on melanin synthesis of B16 melanoma cells. Methods : To investigate melanin synthesis of B16 melanoma cells, this research was measured cell survival, tyrosinase activity, melanin synthesis, western blot. Results : Both Gorynju and Soju extract Psoraleae fructus, cell toxicity depended on the density. Tyrosinase activity depended on the density of Gorynju extract Psoraleae fructus and statistic was showed significant(0.5, 1, 2, 3 ${\mu}g/ml$), in a Soju extract Psoraleae fructus, 1 ${\mu}g/ml$ were showed significant. Melanin synthesis was showed significant in a Soju extract Psoraleae fructus(3, 4 ${\mu}g/ml$). Western blot was showed to depend on the density of Gorynju and a Soju extract Psoraleae fructus. Conclusions : In a tyrosinase activity and a melanin synthesis, the intermediate alcohol of Gorynju and a Soju may be suitable to use.

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Identification and Characterization of Nitric Oxide Synthase in Salmonella typhimurium

  • Choi, Don-Woong;Oh, Hye-Young;Hong, Sung-Youl;Han, Jeung-Whan;Lee, Hyang-Woo
    • Archives of Pharmacal Research
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    • v.23 no.4
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    • pp.407-412
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    • 2000
  • The presence of the nitric oxide synthase (NOS) enzyme from Salmonella typhimurium (S. typhimurium) was identified by measuring radiolabeled L-$[^3H]$citrulline and NO, and Western blot analysis. NOS was partially purified by both Mono Q ion exchange and Superose 12HR size exclusion column chromatography, sequentially. The molecular weight of NOS was estimated to be 93.3 kDa by Western blot analysis. The enzyme showed a significant dependency on the typical NOS cofactors; an apparent Km for L-arginine of 34.7 mM and maximum activity between $37^{\circ}C$ and $43^{\circ}C$. The activity was inhibited by NOS inhibitors such as aminoguanidine and $N^{G}$ $N^{G}$-dimethyl-L-arginine. taken together, partially purified NOS in S. typhimurium is assumed to be a different isoform of mammalian NOSs.OSs.

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Study on the antigenicity of Theileria sergenti merozoite in Korean native cattle (한우(韓牛)에 있어서 Theileria sergenti의 항원성(抗原性)에 관(關)한 연구(硏究))

  • Baek, Byeong-kirl;Kim, Byeong-su;Rhee, Jae-ku
    • Korean Journal of Veterinary Research
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    • v.30 no.2
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    • pp.223-229
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    • 1990
  • A splenectomized 5-month-old calf was inoculated with cryopreserved Theileria sergenti infected blood originated from naturally infected Korean native cattle in Chonbuk district. At peak parasitemia (40.1%), blood was collected, washed, lysed and then the T sergenti merozoite was isolated by differential centrifugation. Antigenic profile of isolated T sergenti organism was analized by SDS-PAGE and western blotting techniques. Coomassie blue stained SDS-PAGE gel revealed at least twelve protein bands of approximately 14Kd, 28Kd, 30Kd, 34Kd, 36Kd, 38Kd, 41Kd, 56Kd, 66Kd, 72Kd, 97Kd and 116Kd in the merozoite homogenate. In western blot, although T sergenti antigen recognized by specific anti-T sergenti antibodies demonstrated 28Kd, 30Kd, 38Kd, 56Kd, 58Kd, 66Kd, 97Kd and 116Kd proteins. False positive reactions were also observed in normal bovine serum with T sergenti and normal erythrocytic antigens. Therefore, predominant proteins of T sergenti merozoite antigen were found to be 28Kd, 30Kd, and 41Kd proteins of molecular weights. On going studies we will analyze the relative importance of those antigens for immunity of T sergenti in Korean native cattle.

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Effects of different parts of Angelica gigas Nakai on brain damages and neuronal death in transient middle artery occlusion/reperfusion-induced ischemic rats (허혈성 뇌졸중 흰쥐에서 당귀의 부위에 따른 뇌신경보호효과 비교 연구)

  • Shin, Yong-Joon;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.29 no.6
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    • pp.85-93
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    • 2014
  • Objectives : We compared with the effects of different parts (root head, root body and hairy root) of Angelica gigas Nakai (Angelicae Gigantis Radix, AG) with on middle cerebral artery occlusion(MCAO)-induced ischemic rats, and on LPS-induced inflammatory response in BV2 microglia. Methods : The 30% ethanol and water extracts of different parts of AG were prepared. Each extract (50 and 100 mg/kg) was administrated intraperitoneally once in MCAO-induced ischemic rats. We measured infarction volumes by TTC staining, and investigated the expression of iNOS, Bax, Bcl-2 and caspase-3 by Western blot. BV2 cells were treated with each extract for 30 min, and then stimulated with LPS. The levels of NO was measured by Griess assay. The expression of iNOS, Cox-2 and proinflammatory cytokines ($TNF-{\alpha}$, $IL-1{\beta}$, and IL-6) were determined RT-PCR and Western blot. The phosphorylation of ERK1/2 and JNK MAPK was determined by Western blot. Results : Among different parts of AG, the 30% ethanol and water extracts of hairy root significantly decreased infarction volume in ischemic brains and inhibited the expression of iNOS, bax and caspase-3. The extracts of hairy root significantly inhibited LPS-induced production of NO, $TNF-{\alpha}$ and IL-6 in BV2 cells, and suppressed the expression of iNOS and COX-2. The hairy root extracts attenuated LPS-induced phosphorylation of ERK1/2 and JNK MAPK in BV2 cells. Conclusions : Our results indicate that the root hairy of AG has a good neuroprotective and anti-inflammatory effects in ischemic stroke compared to other parts.

Suppressive effects of Th2 cytokines expression and the signal transduction mechanism in MC/9 mast cells by flavonol derived from Ginkgo biloba leaves (비만세포에서 은행잎 플라보놀에 의한 Th2 Cytokine 발현 및 신호전달 억제 기전 효과)

  • Kwon, Hae-Young;Chung, Kyu-Jin;Cheong, Kwang-Jo
    • Journal of Digital Convergence
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    • v.13 no.8
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    • pp.503-514
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    • 2015
  • The effects of Flavonol contents from Ginkgo biloba leaf on anti-atopy activity have not rarely been verified. This study is to investigate the effects of flavonol on Th2 cytokine production in MC/9 mast cells. For this, flavonol was analyzed by ELISA and Real-time PCR. Analysis results showed that flavonol significantly suppressed production of Th2 cytokines(IL-13, MIP-1a) in a dose dependent manner. The mRNA expression of IL-4, IL-5, IL-13, TNF-a were effectively restrained by Flavonol at the concentration 25,50,$100{\mu}g/m{\ell}$. And decrease of expression of NFAT-1, c-jun protein was confirmed by western blot analysis. These results indicate that flavonol has effects of decreasing the Th2 cytokine production in the MC/9 mast cell causing inhibition of transcription factors such as NFAT-1, c-jun. Thus, we would like to brief that flavonol may have the applicability as therapeutic agent for atopic dermatitis.

Serologic response of normal Korean children to Pneumocystis carrinii as observed by immunoblot (면역이적법에 의한 한국 어린이의 폐포자충에 대한 항체반응 양상)

  • Mun, Hyeong-Nam;Hong, Seong-Tae;Lee, Sun-Hyeong
    • Parasites, Hosts and Diseases
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    • v.33 no.2
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    • pp.101-106
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    • 1995
  • Soluble protein of purified Pneumocvstis cnrinii was prepared from experimentally infected rats. SDS-PAGE of the crude antigen resolved about 20 protein bands from 20 to 200 kDa. Out of thenl, 116 kDa band strongly reacted and 45-55 and 100 kDa bands reacted weakly to the positive reference human serum from U.S.A. Western blot analysis with sera of 130 normal children and 15 newborns in Korea revealed specific IgG antibody reaction to 40-55 and 116 kDa protein bands. Forty percent (40.0%) of the 145 sera were positive with any of the antigenic protein bands of R corinii. The positive rate was 56% in 50 males and 33.3% in 48 females. The protein bands 40-55 and 116 kDa from rat P. carinii were confirmed to cross-react with human sera in Korea.

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A STUDY ON THE EXPRESSION OF TRANSFORMING GRO WITH FACTOR-β AND MATRIX METALLOPROTEINASE-1 IN PERIAPICAL LESION (치근단질환에서 형질전환성장인자-β와 기질금속함유단백분해효소 발현에 관한 연구)

  • Chi, Jung-Ho;Lee, Su-Jong
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.200-211
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    • 1999
  • The periapical response to injury is a complex interaction of inflammatory, immune, neural, vascular and synthetic activity. TGF-${\beta}$ is a potent modulator of proliferation and differentiation in various tissue, seems to lead to an increase in extracellular matrix. MMP are a family of proteolytic enzyme that mediate the degradation of extracellular matric macromolecules, but little is known about theirs possible role in periapical tissue. The purpose of this study is to investigate the differential expression of TGF-${\beta}$ and MMP-1 in tooth follicle, periapical abscess, granuloma and cyst. The expression of TGF-${\beta}$ and MMP-1 in Periapical tissue was evaluated by immunohistochemical staining and Western blot analysis. Correlationship among the periapical lesions were stastically analyzed. The degree of MMP-1 expression in periapical abscess was higher than in any other periapical lesion, and stastically significant. TGF-${\beta}$ expression is the prominent in granuloma than other periapical lesion, which was stastically significant. The increased expression of MMP and TGF-${\beta}$ was not co-related with inflammatory cell infiltration degree of the periapical cyst. The expression degree of MMP and TGF-${\beta}$ was not co-related with periapical abscess and cyst, but expression of MMP and TGF-${\beta}$ showed strong positive co-relationship with periapical granuloma, which was stastically significant. TGF-${\beta}$ expression by Western blot analysis was prominent in granuloma and cyst, and similar to the results by imunohistochemistry. MMP-1 expression is less than TGF-${\beta}$, but there is not extreme difference between periapical lesion. These results suggest that TGF-${\beta}$ and MMP may be involved in tissue remodeling and has an important role in progress or mediation of periapical lesions.

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