• Journal of Plant Biology
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• 제30권2호
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• pp.109-116
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• 1987

Effects of staining, buffer washing and denaturing agents on the transferrability of RNA fractionated on a methylmercury hydroxide-agarose gel to a nitrocellulose membrane were studied. Ethidium bromide staining and ammonium acetate buffer washing inhibited RNA transfer, while 3% HCHO and 0.5 M NaOH treatments stimulated transfer which was negated in the ammonium acetate buffer. Accordingly, maintenance of primary structure of RNA was proved to be essential for transferring RNA from the methylmercury hydroxideagarose gel to the nitrocellulose membrane.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권4호
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• pp.257-262
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• 2003

A straightforward and effective method is presented for immobilizing enzymes on a microchip platform without chemically modifying a micro-channel or technically microfabricating a column reactor and fluid channel network. The proposed method consists of three steps: the reconstitution of a nitrocellulose (NC) membrane on a plane substrate without a channel network, enzyme immobilization on the NC membrane, and the assembly of another substrate with a fabricated channel network. As a result, enzymes can be stably and efficiently immobilized on a microchip. To evaluate the proposed method, two kinds of enzymatic reaction are applied: a sequential two-step reaction by one enzyme, alkaline phosphatase, and a coupled reaction by two enzymes, glucose oxidase and peroxidase, for a glucose assay.

• BMB Reports
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• 제30권5호
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• pp.308-314
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• 1997

We investigated biotinylation of nitrocellulose membrane for immuno-filtration capture assay. In order to enhance the efficiency of biotinylation, nitrocellulose membranes were pretreated with several chemicals for the purpose of suitable protein absorption through surface modification. As a signal generating enzyme, urease was used and the concentration of avidin was optimized for the efficient binding kinetics between urease-biotin in liquid phase and biotinylated membrane in solid phase. For effective biotinylation, bovine serum albumin-biotin complexes could be immobilized at a concentration of $370\;{\mu}g$/stick ($4.4\;cm^2$). Among tested chemicals, polylysine (0.25%) showed a significant effect in biotinylation. Polylysine is thought to enhance surface area by extending unbound residues into solution. Time of treatment over 30 min and higher molecular weight of polylysines (58,100 dalton) showed positive effect on the enhancement of biotinylation. The result from this study may be useful for developing a new biosensor and other biofunctional membranes for examining molecular recognition.

• KSBB Journal
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• 제11권2호
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• pp.140-150
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• 1996

가정용 혈중 cholesterol 측정시스템을 개발하고자, 그 성능을 조절할 수 었는 벤수들의 최적화가 수 행되었다. 시스템의 주요 구성성분은 분석에 요구되는 효소들 (cholesterol esterase, cholesterol oxidase, 그리고 horseradish peroxidase) 이 부동화된 nitrocellulose membrane strip과 비 이온세척제 (Tri­t ton X-1OO) 및 발색물질 (3,3'-diaminobenzidine) 이 포함된 시료운반용액이다. 시료와 운반용액이 훈 합된 수용액을 membrane의 하단으로부터 흡수시키 면 모세관현상에 의해 시료는 부통화된 효소층으로 전달되어 연속효소반응이 일어난다. 마지막 효소반 응에서 발색물질은 산화되고 그에 따라 cholesterol 농도에 비례한 특정색이 신호로써 발생된다. 그 신호세기에 대해 효소들의 부동화조건과 운반용액의 화학조성이 최적화되었다. 최적조건 하에서 구성된 측정시스템의 농도응탑곡선은 혈중 cholesterol 농도를 측정하기에 충분히 높은 민감도를 나타내었다.

• 농업과학연구
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• 제39권4호
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• pp.613-618
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• 2012

This study was performed to optimize the basic platform of a lateral flow immunoassay. Improvement of the limit of detection (LOD) was evaluated according to the width of a nitrocellulose membrane with varying concentrations of analyte. The analyte, neutravidin was detected based on the avidin-biotin interaction. The antibody-Au nanoparticle conjugation was mostly stabled in a PBS buffer of pH 7.3. The optimal widths of a nitrocellulose membrane were 4 and 6 mm considering the sample flow rate and signal strength of the test line on the membrane. The LOD of neutravidin was 0.001 mg/ml in the optimum conditions.

• KSBB Journal
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• 제11권4호
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• pp.420-430
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• 1996

열처리된 Salmonella typhimurium 항원을 모델 분석물질로 선택하여 단지 시료만을 첨가함으로써 측정을 수행할 수 있는 1단계 immuno--chromatogra phy 분석시스템이 개발되었다. 개발된 시스템은 분 석물질 특정항체(감지항체)-gold 중합체가 건조된 상태로 축적된 glass fiber membrane(하단), 분석 물질의 다른 epltope를 인지하는 항체(포획항체)와 항 감지항체 특정항체들이 공간적으로 분리되어 고정화된 nitrocellulose membrane(중단) 그리고 흡수대로 선택된 cellulose membrane ( 상단)들로 구성된다. 이와 같은 구성성분들은 분석물질을 포함한 시료수용액이 모세관현상에 의해 연속적으로 이동되도록 부분척으로 포개어 배열되었다. 분석시스템의 성능을 조절하는 변수들은 포획항체의 농도 및 membrane 상의 고정화 위치, membrane 표면처리 와 시료운반용액에 샤용된 비반응성 단백질의 종류, 그리고 중합체의 농도로써 확인되었다. 최적조건하 에서, 시스템의 하부로부터 시료수용액을 흡수 후 15분 이내에 포획항체가 고정화된 membrane 상의 지역에 샌드위치 형상의 항원 항체 면역결합체가 형성되었고 이로부터 발생된 발색신호는 분석물질의 농도에 비례하는 것으로 나타났다. 분석물질의 측정 하한농도는 $1{\times}106$ Salmonella cells/mL인 것으로 나타났다.

• The Plant Pathology Journal
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• 제23권4호
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• pp.260-265
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• 2007

Testing a large number of samples from field monitoring and routine indexing is cumbersome and the available virus detection tools were labor intensive and expensive. To circumvent these problems we established tissue blot immunoassay (TBIA) method an alternative detection tool to detect Barley mild mosaic virus (BaMMV) and Barley yellow mosaic virus (BaYMV) infection in the field and greenhouse inoculated plants for monitoring and routine indexing applications, respectively. Initially, leaf and stem were tested to determine suitable plant tissue for direct blotting on nitrocellulose membrane. The dilutions of antibodies were optimized for more efficient and economical purposes. Results showed that stem tissue was more suitable for direct blotting for it had no background that interferes in the reaction. Therefore, this technique was referred as direct stem blot immunoassay or DSBIA, in this study. Re-used diluted (1:1000) antiserum and conjugate up to 3 times with the addition of half strength amount of concentrated antibodies was more effective in detecting the virus. The virus blotted on the nitrocellulose membrane from stem tissues kept at room temperature for 3 days were still detectable. The efficiency of DSBIA and RT-PCR in detecting BaMMV and BaYMV were relatively comparable. Results further proved that DSBIA is a rapid, reliable and economical detection method suitable for monitoring BaMMV and BaYMV infection in the field and practical method in indexing large scale of barley materials for virus resistance screening.

• 센서학회지
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• 제23권6호
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• pp.388-391
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• 2014

The most promising application of the biofuel cells is implantable devices, so the biofuel cells should have an appropriate shape for the vascular vessel. We demonstrated the biofuel cell roll for using in tubes. MWNTs were aggregated by vacuum filtration on a nitrocellulose membrane filter, which was biocompatible and flexible. The MWNT aggregated nitrocellulose membrane used the electrodes of the biofuel cells because it was conductive as well as nanostuructured. Then, the membrane was rolled into the roll shape. The maximum power density of the biofuel cell roll was $7.9{\mu}W/cm^2$ at 153mV and 50 mM glucose. Also, the power density is expected to increase in its practical application if there is flow in the tube, which makes the transportation of fuel easy. The biofuel cell roll contacts with the wall of the tube, so flow in the tube does not disturb. Also, the biofuel cell roll has multi-layers offering more electroactive area.

• Plant Biotechnology Reports
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• 제2권3호
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• pp.171-177
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• 2008

This paper reports an improved protocol for isolation, culture and regeneration of Lotus corniculatus protoplasts. A range of parameters which influence the isolation of L. corniculatus protoplasts were investigated, i.e., enzyme combination, tissue type, incubation period and osmolarity level. Of three enzyme combinations tested, the highest yield of viable protoplasts was achieved with the combination of 2% Cellulase Onozuka RS, 1% Macerozyme R-10, 0.5% Driselase and 0.2% Pectolyase. The use of etiolated cotyledon tissue as a source for protoplast isolation proved vital in obtaining substantially higher protoplast yields than previously reported. Culture of the protoplasts on a nitrocellulose membrane with a Lolium perenne feeder-layer on the sequential series of PEL medium was highly successful in the formation of microcolonies with plating efficiencies 3-10 times greater than previous studies. Shoot regeneration and intact plants were achieved from 46% of protoplast-derived cell colonies.

• Journal of Oral Medicine and Pain
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• 제24권3호
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• pp.235-244
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• 1999

The present study was performed to investigate the binding between salivary proteins(low-molecular-weight mucin;MG2, amylase, proline-rich proteins;PRPs) and oral pathogens(Streptococcus gordonii, Actinomyces viscosus, Staphylococcus aureus) by using solid-phase assay. In the case of transferring proteins to Immobilon-P, S. gordonii binds to MG2. A. viscosus binds to MG2, amylase, and PRPs, and S. aureus binds to MG2 and amylase. On nitrocellulose membrane, S, gordonii and A. viscosus bind to MG2, amylase, and PRPs. S. aureus binds to MG2 and PRPs. However, rabbit anti-A. viscosus antisera and rabbit anti-S. aureus antisera showed cross reactivity to PRPs adsorbed to only nitrocellulose membrane in negative control experiments, which were done without bacterial overlay. The results were different according to the membrane used as solid-phase, which reflected the assay-sensitive nature of binding experiment. PRPs and amylase are known to be components of tooth enamel pellicle. In addition, there was experimental evidence that PRPs and MG2 may covalently bind to oral mucosal epithelium. Considering above facts, the results of the present study can provide information on the interactions between salivary proteins and oral bacteria on tooth and oral mucosal surfaces.