• Journal of Plant Biology
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• v.30 no.2
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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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• v.8 no.4
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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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• v.30 no.5
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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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• v.11 no.2
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• pp.140-150
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• 1996

To develop a home-version assay system for plasma lipoprotein cholesterol, variables that can control the assay performance were optimized. The system was constructcd by using two major components: nitrocellulose membrane strip with immobilized enzymes (cholesterol esterase, cholesterol oxidase, and horseradish peroxidase); and sample carrier solution containing non-ionic detergent (Triton X-100) and chromogen (3,3'-diaminobenzidine). Once a sample combined with the carrier was absorbed from the bottom of the strip, cholesterol was delivered by capillary action to the immobilized enzymes and a sequential reactions took place. In the final reaction, the chromogen was oxidized and then generated a color as signal that was proportional to the concentration of cholesterol. The signal intensity was enhanced by optimizing conditions for the immobilization of enzymes and the chemical composition of carriel. Under these conditions, a dose-response curve was obtained and revealed a high sensitivity enough to measure the cholesterol in blood.

• Korean Journal of Agricultural Science
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• v.39 no.4
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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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• v.11 no.4
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• pp.420-430
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• 1996

One-step immuno-chromatography assay system for heat-killed Salmonella typhimurium antigens was developed. Three major components used were a glass fiber membrane (placed at the bottom of the system) with an antibody (specific to the analyse, detection antibody)-gold conjugate deposited in a dry state on the surface, a nitrocellulose membrane (middle) with an antibody (also, specific to the analyse but recognized different epitome: capture antibody) and anti-detection antibody immobilized in spatially separated areas, and a cellulose membrane (top) as absorption pad. These membranes were partially superimposed such that a wicking of aqueous solution containing sample can continuously take place through membranes. Variables that affected the system performance were the concentration of capture antibody, the location on the membrane, inert protein used for blocking of the membrane and for carrying the sample, and the concentration of the gold conjugate. Under optimal conditions, within 15 minutes after absorption of a sample solution from the bottom of the system antigen-antibody complexes of sandwich type were formed on the membrane surface area with immobilized capture antibody and a color signal was generated in proportion to the analyse concentration. The minimum do tection limit of the analyse was $1{\times}106$ Salmonella cells/mL.

• The Plant Pathology Journal
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• v.23 no.4
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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.

• Journal of Sensor Science and Technology
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• v.23 no.6
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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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• v.2 no.3
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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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• v.24 no.3
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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.