• Title/Summary/Keyword: Protein-A gold

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A Study on the Distribution of Cytochrome-c-oxidase Subunit in the Cristae of Mitochondria (미토콘드리아 크리스테에 존재하는 cytochrome-c-oxidase의 단백질 소단위 분포에 관한 연구)

  • Kim, Soo-Jin;Lee, Ji-Hyon;Chung, Cha-Kwon
    • Applied Microscopy
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    • v.24 no.4
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    • pp.41-51
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    • 1994
  • The topology of the enzyme has been investigated by biochemical studies including chemical labeling and cross linking. Thirteen subunits(polypeptides) of the cytochrome-c-oxidase have localistic characteristics of existing in the matrix side or cytoplasmic side in the mitochondria. In order to observe the distribution of the enzyme subunit on the mitochondria membrane, immunogold-labeling methods were employed. Antibody was obtained from the serum of immunized rabbit with enzyme subunit antigen which was obtained from cytochrome-c-oxidase of the beef heart muscle mitochondria. Beef heart muscle tissue as a tissue antigen was stained with immunized rabbit IgG and protein A gold complex. Electron microscopy has identified the existance of cytochrome-c-oxidase subunit $Mt_I,\;Mt_{II}\;and\;Mt_{III}$ on the membrane of cristae and outer chamber of mitochondria and the subunit $C_{IV}$ on the membrane of cristae and matrix of mitochondria. Particularly, the subunit $C_{IV}$ was also observed to exist in the sarcoplasm of muscle tissue.

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Measurement of Antibodies to Varicella-Zoster Virus Using a Virus-Free Fluorescent-Antibody-to-Membrane-Antigen (FAMA) Test

  • Park, Rackhyun;Hwang, Ji Young;Lee, Kang Il;Namkoong, Sim;Choi, Seuk-Keun;Park, Songyong;Park, Hosun;Park, Junsoo
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.268-273
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    • 2015
  • The fluorescent-antibody-to-membrane-antigen (FAMA) test is regarded as the "gold standard" to detect protective antibodies to varicella-zoster virus (VZV) because of its high sensitivity and specificity. Because the classic FAMA test uses an infectious virus for detection of antibodies to VZV, it is labor-intensive, and also requires special equipment for handling the virus. For this reason, we attempted to develop a simple and safe FAMA assay. Because VZV glycoprotein E (gE) is one of the major VZV glycoproteins, we used the gE protein for the FAMA test (gE FAMA). Here, we demonstrate that overexpression of gE in HEK293T cells can be used to measure antibodies in human serum, and that gE FAMA titers are closely correlated with gpEIA ELISA data. These results indicate that our gE FAMA test has the potential to measure antibodies to VZV.

Immunohistochemical electron microscopic studies on somatotropes and mammotropes in hypophysis of Korean native goat (한국재래산양 뇌하수체의 성장자극세포와 젖샘자극세포에 관한 전자현미경적 연구)

  • Lee, In-se;Lee, Heungshik S.;Won, Moo-ho;Seo, Jehoon;Song, Seung-hoon;Nam, Young-Sam;Kang, Tae-Cheon
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.488-496
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    • 1998
  • Somatotropes, mammotropes and somatomammotropes of the Korean native goat hypophysis were studied by double immunoelectron microscopy using antisera to growth hormone(GH) and prolactin(PRL), and protein A-gold particles of different sizes. Mammotropes were round or oval in shape, and contained round and electron dense secretory granules. The size of secretory granules was variable from 460nm to 680nm in diameter. Somatotropes were elliptical or triangular in shape and the oval nucei were located eccentrically at the periphery of the cell. Secretory granules of the cell were oval in shape and clearly distinguished from round granules of mammotropes. The size of granules was 320~680nm in diameter, smaller than that of mammotropes. Somatomammotropes contained round or oval secretory granules. The granules had intermediate size between somatotropes and mammotropes. Some of granules contained both GH and PRL, while the others contained only one of them.

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Comparison of Growth Characteristics and Productivity of Summer Forage Crops in Sihwa Reclaimed Land (시화 간척지에서 하계 사료작물의 초종 및 품종에 따른 생육특성 및 생산성 비교)

  • Kim, Jong Geun;Jeong, Eun Chan;Kim, Meing Joong;Li, Yan Fen;Kim, Hak Jin;Lee, Su Hwan
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.41 no.2
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    • pp.110-118
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    • 2021
  • This study was conducted to investigate the effects of species and varieties of summer forage crops on growth characteristics and productivity in Sihwa reclaimed land. The summer forage crops used in the trial were silage corn, sorghum×sudangrass hybrid(SSH), and proso millet. For each forage species, Gwangpyeongok(GPO), P15453, P1952 and P2088 were used for silage corn, and 877F, Green star, Honey chew, and Turbo gold cultivars were used for SSH. For proso millet, Ibaekchal, Geumsilchal and Manhongchal developed by the National Institute of Crop Science were used. Silage corn and SSH were sown on May 21, 2019 and proso millet on June 4, and harvested on September 2. There was no significant difference in plant and ear height of silage corn among varieties. P1543 was the highest and P2088 was the lowest in yield of silage corn, but there was no significant difference among treatments. Among the SSH, the plant height of 877F was the highest and Turbo gold variety had the smallest (p<0.05). As for the dry matter(DM) yields, 877F had the highest at 3,862 kg/ha and Green star had the lowest at 2,669 kg/ha (p<0.05). The fresh matter yield of proso millet was 15,778 kg/ha, which was higher than that of corn or SSH, The average dry matter yield was 4,780 kg/ha, and Ibaekchal variety had the highest DM yield compared to other varieties (p<0.05). P2088 had the highest TDN content and GPO was the lowest (p<0.05). As for the SSH, the TDN content of Green star and Honey chew varieties was significantly higher, and the RFV value was the lowest in Turbo gold. The average crude protein content of proso millet was 7.03%, and the highest TDN and RFV values were 64.36% and 106 in Geumsilchal. In the experiment of the germination rate of summer forage crops according to salt concentration, silage corn showed a germination rate of 83.1% even at 0.4% salinity. In particular, P2088 and P1921 varieties had more than 80% germination rate even at 0.6% salt concentration. As for the SSH, the germination rate of 877F was 93.3% even at 0.8% salinity, and 88.3% with Honey chew, indicating higher resistance to salt concentration compared to other varieties. Proso millet showed a high germination rate of 84.0 to 88.7% even at a salt concentration of 0.6%. Considering the above results, proso millet was recommended as the most suitable forage crop species in the Sihwa reclaimed land with high salt concentration, and the Ibaekchal variety is recommended as a suitable forage crop due to its high yield.

Development of Voltammetric Nanobio-incorporated Analytical Method for Protein Biomarker Specific to Early Diagnosis of Lung Cancer (폐암 조기 진단을 위한 단백질 바이오마커 측정용 전압-전류법 기반의 나노바이오 분석법 개발)

  • Li, Jingjing;Si, Yunpei;Nde, Dieudonne Tanue;Lee, Hye Jin
    • Applied Chemistry for Engineering
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    • v.32 no.4
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    • pp.461-466
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    • 2021
  • In this article, a portable and cost-effective voltammetric biosensor with nanoparticles was developed for the measurements of heterogeneous nuclear ribonucleoprotein A1 protein (hnRNP A1) biomarker which can potentially be used for lung cancer diagnosis. Gold nanoparticles were first electrodeposited onto screen printed carbon electrode (SPCE) followed by immobilizing a single stranded DNA aptamer specific to hnRNP A1 onto the electrode surface. Ethanolamine was also used when immobilizing DNA aptamer on the surface to prevent signals from non-specific adsorption events. Sequential injection of hnRNP A1 biomarker and anti-hnRNP A1 conjugated with alkaline phosphatase (ALP) onto the aptamer chip surface allows to form the sandwich complex of DNA aptamer/hnRNP A1/ALP-anti-hnRNP A1 on the electrode surface which further reacted with 4-aminophenyl phosphate (APP). The electrocatalytic reaction of the enzyme, ALP, and the substrate, APP, resulting in the oxidative current response changes at -0.05 and -0.17 V (vs. Ag/AgCl) against the hnRNP A1 concentration was measured using cyclic and differential pulse voltammetry, respectively. The Au nanoparticles-integrated voltammetric biosensor was applied to analyze human normal serum solutions possibly suggesting potential applicability for lung cancer diagnosis.

CHEMICAL AND MICROBIOLOGICAL ANALYSIS OF GOAT MILK, CHEESE AND WHEY BY NIRS

  • Perez Marin, M.D.;Garrido Varo, A.;Serradilla, J.M.;Nunez, N.;Ares, J.L.;Sanchez, J.
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1513-1513
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    • 2001
  • Present Food Legislation compels dairy industry to carry out analyses in order to guarantee the food safety and quality of products. Furthermore, in many cases industry pays milk according to bacteriological or/and nutritional quality. In order to do these analyses, several expensive instruments are needed (Milkoscan, Fossomatic, Bactoscan). NIRS technology Provides a unique instrument to deal with all analytical requirements. It offers as main advantages its speed and, specially, its versatility, since not only allows determine all the parameters required in milk analysis, but also allows analyse other dairy products, like cheese or whey. The objective of this study is to develop NIRS calibration equations to predict several quality parameters in goat milk, cheese and whey. Three sets of 123 milk samples, 190 cheese samples and 109 whey samples, have been analysed in a FOSS NIR Systems 6500 I spectrophotometer equipped with a spinning module. Milk and whey were analysed by folded transmission, using circular cells with gold surface and pathlength of 0.1 m, while intact cheese was analysed by reflectance using standard circular cells. NIRS calibrations were obtained for the prediction of chemical composition in goat milk, for fat (r$^2$=0.92; SECV=0.20%), total solids (r$^2$=0.95: SECV=0.22%), protein (r$^2$=0.94; SECV=0.07%), casein (r$^2$=0.93; SECV=0.07%) and lactose (r$^2$=0.89; SECV=0.05%). Moreover, equations have been performed to determine somatic cells (r$^2$=0.81; SECV=276.89%) and total bacteria (r$^2$=0.58; SECV=499.32%) counts in goat milk. In the case of cheese, calibrations were obtained for the prediction of fat (r$^2$=0.92; SECV=0.57), total solids (r$^2$=0.80; SECV=0.92%) and protein (r$^2$=0.70; SECV=0.63%). In whey, fat (r$^2$=0.66; SECV=0.08%), total solids (r$^2$=0.67; SECV=0.19%) and protein (r$^2$=0.76; SECV=0.07%) NIRS equations were obtained. These results proved the viability of NIRS technology to predict chemical and microbiological parameters and somatic cells count in goat milk, as well as chemical composition of goat cheese and whey.

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Antigenic localities in the tissued of Metagonimus yokogawai observed by immunogoldlabeling method (면역황금 표식법을 이용한 요꼬가와흡충의 조직내 항원성 부위에 관한 연구)

  • Ahn, Hyuk;Rim, Han-Jong;Kim, Soo-Jin
    • Parasites, Hosts and Diseases
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    • v.29 no.3
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    • pp.245-258
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    • 1991
  • In order to determine the antigenic localization in the tissues of the adult Metagonimus yokegawai, immunogoldlabeling method was applied using serum immunoglobulins (IgG) of cats which were infected with isolated metacercariae from Plecoglossus altivelis. The sectioned worm tissue was embedded in Lowicryl HM 20 medium and stained with infected serum IgG and protein A gold complex (particle size: 12 nm) , It was observed by electron microscopy at each tissue of the worm. The gold particles were observed on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum. The gold particles were not observed on the basal lamina of the tegument, interstitial matrix of the parenchyma, the muscle tissue and mitochondria of the tegument. The gold particles were specifically labeled in the secretory granules in the vitelline cells. They were also labeled on the lumen of bladder and egg shell. The above findings showed that antigenic materials in the tissue of adult worms were specifically concentrated on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum.

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Fine Structure and Immunogoldlabeling of Crystalline Inclusion Bodies in Mitochondria (미토콘드리아내 결정함유물의 미세구조 및 면액황금표식법)

  • ;;R.A. Capaldi
    • The Korean Journal of Zoology
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    • v.31 no.1
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    • pp.62-70
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    • 1988
  • The distribution of respiratory chain complexes in beef heart and human muscle mitochondria has been explored by immunoeledron microscopy with antibodies made against beef heart mltochondriai proteins in conjundion with protein A cofloidai gold (l2nm particles). The antibodies used were made against NADH-conezyme Q reductase(complex I), ubiquinol-cytochrome-c-oxldoreductase (complex III) and cytochrome-c-oxidase(complex IV). Labeling of bed heart tissue with any of these antihodies gave gold particles randomly distributed along the mitochondrial inner membrane. The labeling of muscle tIssue mitochondria from a patient with a mitochondrial myopathy localized by biochemical analysis to complex III was quantitated and compared with the labeling of human control muscle tissue mitochondria. Four kinds of morphological changes in the mitochondrial fine strudure in the myopathy patient tissue have been found; paracrystalline inclusions consistIng of densely packed multi- lamellar structures, globular crystalline inclusions with high electron density, multilamellar strudure inclusion body(compadly and irregularly arranged concentric whirl shaped cristae)and golbular cyrstalilne inclusions located in the center of the whirl shaped cristae. Compex I and cytochrome-c-oxldase antihodies reacted to the same level in the mitochondria containing the crystalline inclusions and control mitochondria. Antibodies to complex III reacted very poorly to the mitochondria containing the crystalline Inclusions but strongly to control mitchondria. The globular crystalline inclusions in the mitochondria are not reacted antibodies to respiratory chain complexes.

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Effect of cultivar and ascorbic acid on in vitro shoot regeneration and development of bombardment-mediated plastid transformation of tomato (Lycopersicon esculentum) (토마토 재분화 효율 향상 및 엽록체 형질전환 조건)

  • Roh, Kyung-Hee;Lee, Ki-Jong;Park, Jong-Sug;Kim, Jong-Bum;Lee, Seung-Bum;Suh, Seok-Cheol
    • Journal of Plant Biotechnology
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    • v.37 no.1
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    • pp.77-83
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    • 2010
  • Eighteen cultivars of tomato were tested for their regeneration response. Lycopersicon esculentum cv. 2001-58 showed a very high frequency of regeneration (93%). We evaluated the effect of two compounds with known antioxidant activity (ascorbic acid and cystein). The use of ascorbic acid ($200\;-\;300\;{\mu}M/L$) has a positive effect on shoot regeneration. To develope a system for plastid transformation in tomato via homologous recombination, we constructed the tomato plastid expression vector (pKRT22-AG) harboring 2.2 kb flanking sequences cloned from intact plastid genome and gfp gene. To investigate the factors affecting the delivery system of the pKRT22-AG into chloroplast using bombardment, We assessed the optimal DNA concentration, gold particle volume and target distance. Expression of the GFP protein was observed within chloroplast on protoplast of cotyledon explant by confocal laser scanning microscopy, which indicates that the protocol developed in this study be useful for the production of plastid transgenic plants in tomato.

Immunoelectron Microscopic Observation on the Stratified Squamous Epithelial Cello of Pemphigus vulgaris (Pemphigus vulgaris의 중층 편평상피세포에 대한 면역전자현미경적 관찰)

  • Lee, Cha-Soo;Picut, Catherine-A.;Wilkinson, J.-Erby;Lewis, Robert-M.
    • Applied Microscopy
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    • v.17 no.2
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    • pp.1-22
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    • 1987
  • Pemphigus vulgar교의 본태(本態)를 알아보기 위한 일환으로 본질병(本疾病)의 병변조직(病變組織)을 전자현미경적(電子顯微鏡的)으로 관찰(觀察)하고 본질병(本疾病)에 관여(關與)하는 세포(細胞)들에 대(對)해 면역전자현미경적(免疫電子顯微鏡的)으로 추구(追究)하였던 바, 그 결과(結果)를 보고(報告)하는 바이다. 견(犬)의 pemphigus vulgaris의 구강(口腔) 및 식도(食道)의 점막층(粘膜層)을 전자현미경적(電子顯微鏡的)으로 관찰(觀察)하였던 바 acantholysis를 일으켜도 desmosome과 기저막(基底膜)은 큰 변화(變化)가 없었으며 세포간(細胞間)은 세포간물질(細胞間物質)의 집적(集積)에 의(依)해 확장(擴張)되고 이들 세포간강물질(細胞間腔物質)은 집괴(集塊) 또는 무구조(無構造)한 물질(物質)로 나타났다. 그리고 기저세포(基底細胞)는 기저막(基底膜)에 단단히 부착(附着)되어 있었고 dendritic cell이 기저세포층(基底細胞層)위로 분포(分布)되어 있었으며, 이들 dendritic cell 중(中)에서는 가끔 여러 형태(形態)의 퇴행성변화(退行性變化)를 볼 수 있었다. Mouse 피부유리상피세포(皮膚遊離上皮細胞)에 있어서 immunogold-labeling 방법(方法)에 의해 dendritic cell을 동정(同定)하는 데에는 post-fixation, pre-embedding immunogold-labeling technique가 좋았으며 15nm와 40nm 크기의 colloid-fold 입자(粒子)로 Langerhans cell과 Thy-1양성(陽性) dendritic cell이 표식(標識)될 수 있었다. 이들 세포(細胞)들은 세포막항원(細胞膜抗元)에 따라 monoclonal antibody의 반응(反應)에 이어 치밀한 colloid-gold 입자(粒子)가 세포막표면(細胞膜表面)을 따라 일정(一定)하게 표식(標識)되었다. 또한 이들 상피세포(上皮細胞)들을 투과전자현미경적(透過電子顯微鏡的)으로 관찰(觀察)하였을 때 초미세구조(超微細構造)가 잘 보존(保存)되었으나 Langer-hans cell내(內)의 Birbeck granule은 유리전(遊離前) 피부상피조직내(皮膚上皮組織內)의 Langerhans cell내(內)의 Birbeck granule에 비(比)해 수적(數的)으로 현저히 감소(減少)되어 있었다. 그러나 Thy-1 양성(陽性) dendritic cell에서 볼 수 있는 dense-core 과립(顆粒)은 별변화(別變化)없이 쉽게 관찰(觀察)될 수 있었다. 조직배양(組織培養)을 한 견(犬)의 keratinocyte에 대(對)해 사람 pemphigus vulgaris의 항체(抗體)로 반응(反應)시킨 후 protein-A gold(15 nm)로 표식(標識)시킨 바 제일 바깥 상층(上層)의 keratinocyte에 있어서 세포막표면(細胞膜表面)을 따라 표식(標識)되어 세포막항원(細胞膜抗元)을 나타내었으며, 이와 같은 소견(所見)으로 미루어 정상피부(正常皮膚) 중층편평상피세포(重層扁平上皮細胞)에서도 동일(同一)한 소견(所見)을 관찰(觀察)할 수 있다고 본다.

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